Indian Journal of Experimental Biology

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VOLUME 51

NUMBER 11

NOVEMBER 2013

CODEN: IJEB (A6) 51 (11) 773-866 (2013)

ISSN: 0019-5189 (Print); 0975-1009 (Online)

CONTENTS

Special Issue on New Developments in Biotechnology 

Preface

869

 

 

Review Articles

 

 

 

Production, purification, characterization and over-expression of xylanases from actinomycetes

875

 

 

Leya Thomas, Abhilash Joseph, Muthu Arumugam & Ashok Pandey

 

 

 

Molecular regulation of cholesterol metabolism: HDL-based intervention through drugs and diet

885

 

 

Amit Kumar Rai, Patrizia Debetto & Federica Dabbeni Sala

 

 

 

Relevance of physical properties in the stability of plant-based food products

895

 

 

Elena Venir & Enrico Maltini

 

 

 

Mini Review

 

 

 

Survival strategies of Bacillus spores in food

905

 

 

Mara Lucia Stecchini, Manuela Del Torre & Pierluigi Polese

 

 

 

Papers

 

 

 

Potentialities of newly isolated Bacillus subtilis and Lactobacillus sp for curd preparation
and a comparative study of their physico-chemical parameters with other marketed curds

910

 

 

Atanu Adak, Saswati Parua (Mondal), Chiranjit Maity, Kuntal Ghosh, Suman K Halder,
Pradeep K Das Mohapatra & Keshab C Mondal

 

 

 

In vitro propagation of a rare medicinal fern of Western Ghats–Diplazium esculentum (Reytz.)

919

 

 

Archana G Nair,  S Pradeesh, G S Nikhila, G Sangeetha, I Mini & T S Swapna

 

 

 

Exploitation of fermented shrimp-shells hydrolysate as functional food: Assessment of antioxidant, hypocholesterolemic and prebiotic activities

924

 

 

Suman Kumar Halder, Atanu Adak, Chiranjit Maity, Arijit Jana, Arpan Das, Tanmay Paul,
Kuntal Ghosh, Pradeep Kumar Das Mohapatra, Bikas Ranjan Pati & Keshab Chandra Mondal

 

 

 

Statistical media and process optimization for biotransformation of rice bran to vanillin
using Pediococcus acidilactici,

935

 

 

Baljinder Kaur & Debkumar Chakraborty

 

 

 

Mathematical model-based optimization of physico-enzymatic hydrolysis of Pinus roxburghii needles for the production of reducing sugars

944

 

 

Siddharth Vats, Devendra Prasad Maurya, Ayushi Jain, Varija Mall & Sangeeta Negi

 

 

 

Low cost single-step purification of endoglucanase from Aspergillus fumigatus ABK-9

954

 

 

Arpan Das, Tanmay Paul, Suman K Halder, Arijit Jana, Kuntal Ghosh, Chiranjit Maity,
Pradeep K Das Mohapatra, Bikas R Pati & Keshab C Mondal

 

 

 

Analysis of alteration of gut microbial population under the exposure of graded hyperbaric pressures: Application of metagenomic approach

960

 

 

Chiranjit Maity, Atanu Adak, Suman Kumar Halder, Arijit Jana, Kuntal Ghosh,
Pradeep Kumar Das Mohapatra, Bikas Ranjan Pati & Keshab Chandra Mondal

 

 

 

Biosynthesis of colloidal gold nanoparticles by Streptomyces sp. NK52 and its anti-lipid
peroxidation activity

969

 

 

Divya Prakash, Vishal Mahale, Ashok Bankar, Neelu Nawani, Smita Zinjarde &
Balasaheb Kapadnis

 

 

 

Biosynthesis and characterization of mercury sulphide nanoparticles produced by
Bacillus cereus MRS-1

973

 

 

S Sathyavathi , A Manjula, J Rajendhran J & P Gunasekaran

 

 

 

Optimization of lipid enriched biomass production from oleaginous fungus using response
surface methodology

979

 

 

S P Jeevan Kumar & Rintu Banerjee

 

 

 

Characterization and antimicrobial activity of lectins from Penicillium sp.

984

 

 

RS Singh, P Jain & HP Kaur

 

 

 

Optimization of enzymatic saccharification of microwave pre-treated sugarcane tops through response surface methodology for biofuel

992

 

 

Devendra Prasad Maurya, Siddharth Vats , Sudheer Rai & Sangeeta Negi

 

 

 

A comparative study on cell disruption methods for release of aspartase from E. coli K-12

997

 

 

RS Singh

 

 

 

Isolation of a lead tolerant novel bacterial species, Achromobacter Sp. TL-3: Assessment of bioflocculant activity

1004

 

 

Neha Batta, Sanjukta Subudhi, Banwari Lal & Arundhuti Devi

 

 

 

Evaluation of polymeric adsorbent resins for efficient detoxification of liquor generated
during acid pretreatment of lignocellulosic biomass

1012

 

 

Soolamkandath Variem Sandhya, Kumar Kiran, Mathiyazhakan Kuttiraja, Varghese Elizabeth Preeti, Raveendran Sindhu, Sankar Vani, Sukumaran Rajeev Kumar, Ashok Pandey & Parameswaran Binod

 

 

 

Treatment of waste gas containing low concentration of dimethyl sulphide in a high performance biotrickling filter

1018

 

 

Balendu Shekher Giri, Asha Juwarkar, S N Mudliar & R A Pandey

 

 

 

Proteolytic enzyme mediated antagonistic potential of Pseudomonas aeruginosa against Macrophomina phaseolina

1024

 

 

Devaraj Illakkiam, Nishanth Lipton Anuj, Paramasivan Ponraj, Manoharan Shankar,
Jeyaprakash Rajendhran & Paramasamy Gunasekaran

 

 

 

Candida albicans biofilm inhibition by synergistic action of terpenes and fluconazole

1032

 

 

Suma C Pemmaraju, Parul A Pruthi, R. Prasad & Vikas Pruthi

 

 

 

Liquid based formulations of bacteriophages for the management of waterborne bacterial pathogens in water microcosms

1038

 

 

Sangeeta Ahiwale, Sujata Tagunde, Sushama Khopkar, Mrudula Karni, Milind Gajbhiye &
Balasaheb Kapadnis

 

 

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Author Index

Adak Atanu

910,924960

Ahiwale Sangeeta

1038

Anuj Nishanth Lipton

1024

Arumugam Muthu

875

 

 

Banerjee Rintu

979

Bankar Ashok

969

Banwari Lal

1004

Batta Neha

1004

Binod Parameswaran

1012

 

 

Chakraborty Debkumar

935

 

 

Das Arpan

924,954

Debetto Patrizia

885

Devi Arundhuti

1004

 

 

Gajbhiye Milind

1038

Ghosh Kuntal

910,924,954,960

Giri Balendu Shekher

1018

Gunasekaran Paramasamy

973,1024

 

 

Halder Suman K

910,924,954,960

 

 

Illakkiam Devaraj

1024

 

 

Jain Ayushi

944

Jain P

984

Jana Arijit

924,954,960

Jeevan Kumar S P

979

Joseph Abhilash

875

Juwarkar Asha

1018

 

 

Kapadnis Balasaheb

969,1038

Karni Mrudula

1038

Kaur Baljinder

935

Kaur H P

984

Khopkar Sushama

1038

Kiran Kumar

1012

Kuttiraja Varghese Elizabeth Preeti

1012

 

 

Mahale Vishal

969

Maity Chiranjit

910,924,954,960

Mall Varija

944

Maltini Enrico

895

Manjula Armugam

973

Mathiyazhakan

1012

Maurya Devendra Prasad

944,992

Mini I

919

Mohapatra Pradeep Kumar Das

910,924,954,960

Mondal Keshab Chandra

910,924,954,960

Mudliar S N

1018

 

 

Nair Archana G

919

Nawani Neelu

969

Negi Sangeeta

944,992

Nikhila G S

919

 

 

Pandey Ashok

875,1012

Pandey R A

1018

Parua (Mondal) Saswati

910

Pati Bikas Ranjan

924,954,960

Paul Tanmay

924,954

Polese Pierluigi

905

Ponraj Paramasivan

1024

Pradeesh S

919

Prakash Divya

969

Prasad R

1032

Pruthi Parul A

1032

Pruthi Vikas

1032

 

 

Rai Amit Kumar

885

Rai Sudheer

992

Rajeev Kumar Sukumaran

1012

Rajendhran Jeyaprakash

973,1024

 

 

Sala Federica Dabbeni

885

Sangeetha G

919

Sankar Vani

1012

Sathyavathi Sundararaju

973

Shankar Manoharan

1024

Sindhu Raveendran

1012

Singh R S

984,997

Stecchini Mara Lucia

905

Subudhi Sanjukta

1004

Suma C Pemmaraju

1032

Swapna T S

919

 

 

Tagunde Sujata

1038

Thomas Leya

875

Torre Manuela Del

905

 

 

Variem Sandhya Soolamkandath

1012

Vats Siddharth

944,992

Venir Elena

895

 

 

Zinjarde Smita

969

 

 

Keyword Index

7-11 phages

1038

9-Octadecenoic acid methyl ester

979

 

 

Achoromobacter

1004

Actinomycetes

875

Adsorbing column

954

Aeromonas hydrophila

924

Antifungal therapy

1032

Anti-lipid peroxidation

969

Antimicrobial

910

Antioxidant

910,924

Aspartase

997

Aspergillus fumigatus

954

 

 

Bacillus

905

Bacillus cereus

973

Bacteriophages

1038

Biofilm

1032

Bioflocculant

1004

Biofuel

875

Biomass

875

Biosorption

1004

Biotrickling filter

1018

 

 

C. albicans

1032

Cardiovascular disease

885

Cell disruption

997

Cellulase

944,992

Cholesterol

885

Curd

910

 

 

Dimethyl sulphide

1018

Diplazium esculentum

919

Drug resistant bacterial pathogen

1038

Drugs interfering with RCT

885

Dryopteridaceae

919

 

 

E. coli

997

Environmental bioremediation

973

Enzymatic pre-treatment

944

 

 

Flavonoid

910

Food

905

Functional food

924

 

 

Fungal lectins

984

Furfural

1012

 

 

GC-MS

979

Gel electrophoresis

954

Glass transition

895

 

 

Haemagglutination inhibition

984

HDL

885

Hexadecanoic acid methyl ester

979

HMF

1012

Hyperbaric pressure

960

Hypocholesterolemic

924

 

 

In vitro shooting

919

Intestinal microflora

960

 

 

Kaolin clay

1004

 

 

Laccase

944

Lactic acid bacteria

910

Langmuir adsorption isotherm

954

Lignocellulosic biomass

1012

Lipid

979

Liquid-shear

997

Loading

1018

 

 

Mercury

973

Mercury sulphide nanoparticle

973

Metagenomics

960

Microwave pretreatment

992

Monovalent liquid formulations

1038

Morcophomina phaseolina

1024

MS medium

919

 

 

Nanoparticles

969

Nutrition

885

 

 

Oleaginous fungus

979

Oxidative stress

924

 

 

Pb

1004

PCR-DGGE

960

Pediococcus acidilactici

935

Penicillium corylophilum

984

Penicillium expansum

984

Penicillium purpurogenum

984

Phenotypic variability

905

Plant food

895

Plant growth promoting rhizobacteria

1024

Poly urethane foam

1018

Polyphenol

910

Pretreatment liquor

1012

Protease

1024

Pseudomonas aeruginosa

1024

 

 

Removal efficiency

1018

Rhizosphere

1024

Rice bran

935

RSM

935,944,979

 

 

Saccharification

992

Shrimp-shells hydrolysate

924

Single-step purification

954

Solid-shear

997

Sonication

997

Spore

905

Stability

895

Statistical optimization

935

Steam explosion

944

Streptomyces

969

Sugarcane top

992

Synergy

1032

 

 

T7-like

1038

Terpenes

1032

Tolerance

1004

 

 

Vanillin

935

 

 

Water activity

895

 

 

XAD

1012

Xylanase

875,944

 

 

Zymogram

954

 

 

 

 

Preface

 

Industrial biotechnology, often also referred as white biotechnology, involves the utilization
of modern biotechnology for the production of industrial commodity and fine chemicals
and products such as enzymes, biopolymers, organic acids, vitamins, biofuels, biofertilizers,
biopesticides, etc. Such processes use simple (such as glucose and sucrose), or complex
(such as molasses, starches, etc) polysaccharides as the source of carbon source for cultivating
the microbes. These bioprocesses could also involve bioconversion or biotransformation of
organic molecules using enzyme to produce the desirable products. The domain of industrial
bioprocessing is widely regarded as the third wave of biotechnology, distinct from the first
two waves, i.e., medical, or red biotechnology, and agricultural, or green biotechnology.
In biotechnological processes, microbial, animal/plant cells, or enzymes are used to make
the products for various industrial sectors, including chemicals, pharmaceuticals, food & feed, detergents, pulp & paper, textiles, energy, materials & polymers, etc.

The use of microorganisms to transform the biological materials into fermented food
products has its origins since long and is regarded the oldest food processing technology.
With the advent of biotechnology, bioprocesses have been developed for an enormous range
of commercial products, from relatively cheap materials, such as industrial alcohol and organic solvents to expensive specialty chemicals such as antibiotics, therapeutic proteins and vaccines. As techniques and instrumentation are refined, bioprocesses find applications in other newer
areas where chemical processes are used conventionally.

The domain of industrial biotechnology has great significance because this offers potential
environmental and economic benefits associated with reduced energy consumption, green-house gases emissions and waste generation. New biotechnology is expected to enable the paradigm shift from fossil fuel-based to bio-based production of value-added chemicals. The fundamental force that drives the development and implementation of industrial biotechnology is the market economy, as biotechnology promises highly efficient processes at lower operating and capital expenditures. In addition, political and societal demands for the sustainability and environment-friendly industrial production systems, coupled with depletion of crude oil reserves and a growing world demand for raw materials and energy will continue to drive this trend
forward. Government policies including tax incentives, mandatory-use regulations, research &
development, commercialization support, loan guarantees and agricultural feedstock support
programme have also greatly helped to the adoption of industrial biotechnology. Perceived needs and marketability, the researcher’s imagination, ethics, and governmental regulations
essentially are the major factors in setting the stage and boundaries for developments in industrial biotechnology. The breakthroughs in enzyme engineering, metabolic engineering, synthetic
biology and the expanding “omics” toolbox coupled with computational systems biology
are expected to speed-up industrial applications of biotechnology. These advances have provided scientists with toolsets to engineer enzymes and whole cells by expanding the means to identify, understand and make perturbations to the complex machinery within the microorganisms.

This special issue of the Indian Journal of Experimental Biology comprises the selected
peer-reviewed papers presented in the International Conference on Industrial Biotechnology (ICIB-2012), the IX Convention of the Biotech Research Society, India (BRSI) and Indo-Italian Workshop on Food Biotechnology: Industrial Processing, Safety & Health. The conference was held at the Punjabi University, Patiala, India, during 21st-23rd November 2012 and focused
on recent developments on the frontier areas of industrial biotechnology as the theme. The conference also covered the highly relevant topic in the areas of Food & Agricultural Biotechnology, Environmental Biotechnology and Medical Biotechnology and brought together a multinational body of scientists and experts to deliberate on global developments in various fields of biotechnology and their applications in industrial, medical, environmental, food and
agriculture sectors. A large number of participants from all over the world attended it.
The conference not only provided a unique platform to the participants from the industries
and academic institutions to share their thoughts and views to pave the way to develop possible linkages among them, but also served the purpose of global networking among them and helped in creating a nucleus of interface research.

The scientific programme of the conference included invited talks from the internationally
eminent experts and poster presentation by the contributory authors. During the three days
deliberations, more than 600 delegates, including 50 overseas delegates from various countries took part in discussions. About 20 delegates from various Biotech industries also participated
in this event. Overall, there were 106 lectures in various fields of Biotechnology presented by
the eminent scientists who shared their research experiences in a total of 24 technical sessions.
There has been huge response from authors for the submission of the manuscripts for publication. This issue contains 23 papers, which have been selected for publication after peer-review.

The organizing committee of the ICIB-2010 would like to thank the invitees and delegates for their overwhelming response for the conference and participating in the discussions actively. Thanks are also due to the session coordinators, session chairs, poster evaluation juries, members of national and international advisory committees and local organizing committee for their
strong efforts and help in smooth conduction of the conference. We gratefully acknowledge
the support and participation of national and international organizations and agencies, which
extended sponsorship to the event. Without their support and participation, it would have not been easy to make suitable arrangements for the conference. The Guest Editors also gratefully
acknowledge the support by the French government research program ‘Investissements d’avenir’ through the IMobS3 Laboratory of Excellence (ANR-10-LABX-16-01), by the European Union through the program Regional competitiveness and employment 2007-2013 (ERDF-Auvergne region) and by the Auvergne region.

We are grateful to the Director, CSIR-National Institute of Science Communication
And Information Resources (NISCAIR), New Delhi and Mr. Rajiv Mathur, Editor of the Indian Journal of Experimental Biology, who graciously agreed to our proposal to bring out this special
issue of the journal based on the peer-reviewed papers. We express our sincere gratitude to the reviewers who evaluated the manuscripts in a timely fashion and are thankful to all others at the Editorial Office of the journal associated with the bringing out this special issue.

 

Guest Editors

Prof. Ram Sarup Singh

Prof. Ashok Pandey

Prof. Claude-Gilles Dussap

Prof Luciano Piergiovanni

Department of
Biotechnology

Punjabi University

Patiala 147 001,  
India

Centre for Biofuels and
Biotechnology Division

CSIR-National Institute
for Interdisciplinary
Science and Technology (NIIST) Trivandrum
695 019, India

 

Polytech Clermont Ferrand

Chemical & Biochemical
Engineering Laboratory

Institute Pascal, Universit Blaise Pascal

Clermont Ferrand, France

Department of Food,
Environmental and
Nutritional Sciences,

University of Milan

Milano, Italy

 

 

 

 

            Correspondent author is marked by *

 

 

Review Articles

 

 

Indian Journal of Experimental Biology

Vol. 51, November 2013, pp. 875-884

 

 

 

Production, purification, characterization and over-expression of xylanases from actinomycetes

Leya Thomas*, Abhilash Joseph1, Muthu Arumugam & Ashok Pandey

Biotechnology Division, CSIR-National Institute for Interdisciplinary Science and Technology
(NIIST), Trivandrum 695 019, India
1Department of Biotechnology & Microbiology, School of Life Science, Kannur University, Kannur 670 661, India

 

Xylanases are a group of depolymerizing enzymes often used for the hydrolysis of xylan (present in hemicellulose) to monomeric sugars and comprise endo-xylanases (EC 3.2.1.8) and β-xylosidases (EC 3.2.1.37). They often act in synergy with other enzymes for complete hydrolysis of hemicellulose. Xylanases find several industrial applications, for example in food and feed industries, paper and pulp industries and more recently have acquired a great role in biomass to biofuels program. Bacteria and fungi can best produce xylanases. Recent developments in rDNA technology have resulted in molecular cloning and expression of xylanases in heterologous and homologous hosts. In view of significance of the actinomycetes for the production of biotechnological products, attempts have been made in recent years to explore them for the production of industrial enzymes, including xylanses, aiming to find the enzyme with novel features. This review provides the state-of-art information and developments on the xylanases from actinomycetes, presenting the production, purification, characterization and over-expression from various actinomycetes cultures.

 

 

Indian Journal of Experimental Biology

Vol. 51, November 2013, pp. 885-894

 

 

Molecular regulation of cholesterol metabolism: HDL-based intervention
through drugs and diet

Amit Kumar Rai, Patrizia Debetto & Federica Dabbeni Sala*

Department of Pharmaceutical and Pharmacological Sciences, University of Padova,
Largo E. Meneghetti, 2, 35131 Padova, Italy

The overloading of cholesterol in the arteries remains the principal cause of cardiovascular diseases. Since available anti-cholesterolemic drugs are not completely effective and have several severe adverse effects, the aim of this review is to analyze current research focused on the emerging, innovative therapeutic strategies based on both pharmacological and nutritional interventions to control cholesterol metabolism. Pharmacological interventions mainly involve the use of molecules capable of interfering with high-density lipoprotien (HDL) metabolism and the reverse cholesterol transport (RCT) through genetic control of apolipoprotein A-I (ApoA-I), agonism at liver X-receptor a (LXRa), or inhibition of cholesteryl ester transport protein (CETP), scavenger receptor BI(SR-BI), and ecto F0F1ATPase/synthase. Nutritional interventions are based on the use of fibres, phytosterols, and probiotics acting through interference with absorption and re-absorption of cholesterol by enterocyte and hepatocyte specific transporters, thus influencing RCT final step. The search for new drugs is still at the very beginning and new molecules are not yet ready to enter clinical use. However, several promising findings coming from innovative biotechnological research are expected shortly to produce probiotics, fibres, and phytosterols to be used as therapeutic tools. Among the most important advantages of natural products in respect to traditional drugs are the lack of severe adverse effects and their low cost.

 

 

Indian Journal of Experimental Biology

Vol. 51, November 2013, pp. 895-904

 

 

Relevance of physical properties in the stability of plant-based food products

Elena Venir* & Enrico Maltini

Department of Food Science, University of Udine, Via Sondrio 2/A, 33100 Udine, Italy

 

Plant tissues are composed of a watery solution of low molecular weight species, mainly sugars, salts and organic acids, and of high molecular weight hydrocolloids, contained in a water insoluble matrix of macromolecules, mostly carbohydrates. All these constituents interact with water, thus reducing its thermodynamic vapour pressure (aw), with small molecules interacting through polar binding, and large biopolymers through surface and capillary effects. Similarly, some constituents will greatly affect kinetic glass transition temperatures (Tg), while others will not. As regards stability, while microbial and chemical changes are mainly related to aw, structure-related changes such as collapse are dependent on the glass transition temperature, Tg. In simple systems such as juices, both thermodynamic and kinetic approaches, employed respectively for high and low moisture systems, have predictive ability, which can be unified in the concept of “critical aw”. However, in complex, multidomain, multiphase systems, such as vegetables and fruits, where insoluble polymeric phases are present, hydrocolloids such as soluble pectins will only slightly affect Tg and aw, but significantly increase the macro viscosity of the soluble fraction, thereby reducing the tendency to collapse. In such cases the use of Tg as a predictive tool must be considered with care. The interrelationships among these aspects are discussed in detail below.

 

 

Indian Journal of Experimental Biology

Vol.51, November 2013, pp. 905-909

 

 

 

Mini Review

Survival strategies of Bacillus spores in food

 

Mara Lucia Stecchini1*, Manuela Del Torre1 & Pierluigi Polese2

1Department of Food Science, via Sondrio 2/a, University of Udine, 33100 Udine, Italy

2Department of Chemistry, Physics and Environment, via del Cotonificio 108, University of Udine, 33100 Udine, Italy

Control of bacterial spores is one of the major problem in the food preservation. Spores of Bacillus genus are commonly present in different environments, including soil and the gut of insects and animals and, as a result, they can be spread to all kind of foods. Due to their high resistance properties, their complete inactivation in food is often impossible without changing the product characteristics. Surviving spores can germinate and grow out to vegetative cells, with the consequent great risk of food spoilage and food poisoning after consumption. Spores have evolved various mechanisms, including phenotypic variability, to protect themselves from a wide range of damage resulting from food preservation treatments. Even if the phenotypic heterogeneity contributes to increase the chances of survival of Bacillus spore to conventional preservation treatments, in some specific instances, an homogeneous response could be the result of a strategy adopted by the spores to increase resistance to those treatments.

 

Papers

 

Indian Journal of Experimental Biology

Vol. 51, November 2013, pp. 910-918

 

 

Potentialities of newly isolated Bacillus subtilis and Lactobacillus sp for curd preparation and a comparative study of its physico-chemical parameters
with other marketed curds

Atanu Adak, Saswati Parua (Mondal), Chiranjit Maity, Kuntal Ghosh, Suman K Halder,
Pradeep K Das Mohapatra & Keshab C Mondal*

Department of Microbiology, Vidyasagar University, Midnapore 721 102, India

Department of Physiology, Bajkul Milani Mahavidyalaya, Purba Medinipur, India

Received 6 December 2012; revised 18 February 2013

Two Bacillus sp. were isolated from the local fermented milk and identified on the basis 16S rRNA sequence profile as Bacillus subtilis AKL1 and by biochemical process as Lactobacillus acidophilus AKL2. These isolates were used as fresh inoculums for curd preparation individually and in combinations. Different physico-chemical and therapeutic properties of the newly prepared curd were examined and compared with marketed local (sweet and sour) and branded (Mother Dairy and Thackar) curds. The total hydrolyzed peptides, free amino acids, lactic acid were significantly higher, whereas, total solid, ash content, syneresis and free reducing sugar were lower in the curd prepared by a mixture of AKL1 and AKL2 (0.5:0.5, v/v). The antioxidant activity against ABTS+, DPPH, OH and Fe3+ were also higher in the newly formulated curd. Polyphenols (85.5µg/g), flavonoids (12.5µg/g) and free aromatic amino acids contents were also higher in AKL1+AKL2. All these components prevent excess protein oxidation that was revealed by SDS-PAGE. The curd also exhibited potent antimicrobial activity against some entero-pathogens like Clostridium perfringens, Escherichia coli, Shigella dysentery, Vibrio cholerae and Staphylococcus aureus. It can be concluded that the combination of these Lactobacillus sp. will be a fruitful inoculum for the preparation of curd having better health promoting effects.

 

 

Indian Journal of Experimental Biology

Vol. 51, November 2013, pp. 919-923

 

 

In vitro propagation of a rare medicinal fern of Western Ghats – Diplazium esculentum (Reytz.)

Archana G Nair, S Pradeesh, G S Nikhila, G Sangeetha, I Mini & T S Swapna *

Department of Botany, University College, Trivandrum 695 034, India

Received 8 December 2012; revised 9 March 2013

Present study aimed for in vitro culture of circinate part of young leaves of D. esculentum which is amongst the
leafy vegetables consumed as vegetable by Paniya and Chetti tribes of Western Ghats. The circinate part of young
leaves (crosiers), excised before the beginning of foliar expansion, was inoculated on half strength Murashige and
Skoog (MS) medium supplemented with auxins indole-3-butyric acid (IBA) or α-napthalene acetic acid (NAA) or
2,4-Dichlorophenoxyacetic acid (2,4-D) and cytokinin 6- benzylaminopurine (BA) in a range 0.5 to 2.5 mg L-1. Combinations of different concentrations of 2,4 D + BA, IBA + BA as well as of NAA+ BA were also tested in half strength MS medium with 3% sucrose and with pH 5.8. The best morphogenic response was obtained with half strength MS medium supplemented with 2,4-D 0.5 mg L-1 and BA 2.5 mg L-1, 3% sucrose, at pH 5.8. For rooting of the microshoots, half strength MS medium supplemented with 2,4-D ( 2 and 1 mg L-1 ) exhibited best results. Present study reports the successful in vitro culturing of D. esculentum.

 

 

Indian Journal of Experimental Biology

Vol. 51, November 2013, pp. 924-934

 

 

Exploitation of fermented shrimp-shells hydrolysate as functional food: Assessment of antioxidant, hypocholesterolemic and prebiotic activities

 

Suman Kumar Halder, Atanu Adak, Chiranjit Maity, Arijit Jana, Arpan Das, Tanmay Paul, Kuntal Ghosh,
Pradeep Kumar Das Mohapatra, Bikas Ranjan Pati* & Keshab Chandra Mondal

Department of Microbiology, Vidyasagar University, Midnapore 721 102, India

Received 9 December 2012; revised 16 February 2013

In the present study the bioactivities of chitooligosaccharides of fermented shrimp-shell hydrolysate (SSH) in respect to hypocholesterolemic, antioxidant and prebiotic activity were tested in male albino rat. Rats were treated with four different diets, viz., (i) cholesterol-rich (5%) basal diet (ChB), (ii) ChB+10% chitin, (iii) ChB+10% SSH and (iv) control group (without cholesterol). After 4 weeks of treatment, body mass index, liver weight, serum total cholesterol and
LDL-cholesterol in groups (ii) and (iii) were decreased significantly than group (i). SSH supplementation significantly resists oxidative stress by reducing the thiobarbituric acid reactive substances and by increasing catalase, superoxide dismutase and free radical scavenging activity. The colonization of Lactobacillus and Bifidobacterium population in small and large intestine were more in group (iii) than other groups. Reduction of Clostridium perfringens population and
non-significant changes of E. coli was also noted in SSH supplement group. Histological study revealed that the villus height and villus:crypt of the small intestine were increased significantly in SSH supplemented group (iii) without any diarrheal symptoms. The results demonstrated that the shrimp-shells hydrolysate has hypocholesterolemic effect, can resist lipid peroxidation and can influence the growth of health beneficial microbes, hence can be used as functional food for hypercholesterolemic patients.

 

 

Indian Journal of Experimental Biology

Vol. 51, November 2013, pp. 935-943

 

 

Statistical media and process optimization for biotransformation of rice bran
to vanillin using Pediococcus acidilactici

Baljinder Kaur* & Debkumar Chakraborty

Department of Biotechnology, Punjabi University, Patiala 147 002, India

Received 7 December 2012; revised 9 March 2013

An isolate of P. acidilactici capable of producing vanillin from rice bran was isolated from a milk product. Response Surface Methodology was employed for statistical media and process optimization for production of biovanillin. Statistical medium optimization was done in two steps involving Placket Burman Design and Central Composite Response Designs. The RSM optimized vanillin production medium consisted of 15% (w/v) rice bran, 0.5% (w/v) peptone, 0.1% (w/v) ammonium nitrate, 0.005% (w/v) ferulic acid, 0.005% (w/v) magnesium sulphate, and 0.1% (v/v) tween-80, pH 5.6, at a temperature of 37 °C under shaking conditions at 180 rpm. 1.269 g/L vanillin was obtained within 24 h of incubation in optimized culture medium. This is the first report indicating such a high vanillin yield obtained during biotransformation of ferulic acid to vanillin using a Pediococcal isolate.

 

 

Indian Journal of Experimental Biology

Vol. 51, November 2013, pp. 944-953

 

 

Mathematical model-based optimization of physico-enzymatic hydrolysis of
Pinus roxburghii needles for the production of reducing sugars

 

Siddharth Vats, Devendra Prasad Maurya, Ayushi Jain, Varija Mall & Sangeeta Negi*

Motilal Nehru National Institute of Technology, Allahabad 211004, India

Received 14 January 2013; revised 11 March 2013

The objective of this study was to optimize the physico-enzymatic pretreatment of P.roxburghii fallen foliage (needles) to produce reducing sugars through response surface methodology (RSM) with central composite face centered design (CCD). Under this, five parameters, i.e., concentration of laccase, cellulose and xylanase, steam explosion pressure and incubation period, at three levels with twenty six runs were taken into account. Cellulase, xylanase and laccase enzymes with activity 4.563, 38.32 and 0.05 IU/mL, respectively, were produced from locally isolated microbial strains. The analysis of variance (ANOVA) was applied for the validation of the predicted model at 95% of confidence level. This model predicted 334 mg/g release of reducing sugars on treating P.roxburghii fallen foliage with 1.18 mL of cellulose, 0.31 mL of xylanase and 0.01 mL of laccase, 14.39 psi steam explosion pressure and 24 h of incubation time. The experimental results obtained were in good agreement to predicted values, making it a reliable optimized model for five factors in combination to predict reducing sugar yield for ethanol production for bio-fuel industry.

 

 

Indian Journal of Experimental Biology

Vol. 51, November 2013, pp. 954-959

 

Low cost single-step purification of endoglucanase from
Aspergillus fumigatus ABK-9

 

Arpan Das, Tanmay Paul, Suman K Halder, Arijit Jana, Kuntal Ghosh, Chiranjit Maity,
Pradeep K Das Mohapatra, Bikas R Pati & Keshab C Mondal*

Department of Microbiology, Vidyasagar University, Midnapore 721 102, India

Received 7 December 2012; revised 2 March 2013

Low cost agro-waste was used as adsorption support for single-step purification of endoglucanase from the culture filtrate of A. fumigatus ABK-9. Among various agro-waste substrates, 1% NaOH pretreated rice bran was proved to be the best for adsorbing about 74.8 and 71.1% of endoglucanase at 4 °C and 10 °C respectively. Langmuir type adsorption isotherm at 4 °C showed maximum adsorption of enzyme at pH 5.0, which was in the range of optimum pH of the enzyme. The rice bran column bound enzyme was maximally eluted by a mixture of acetate buffer (0.05 M, pH 5.5) and ethanol (40%, v/v) at a ratio of 3:2 and a flow rate of 1 mL/min. A 5.52-fold purification of the enzyme was achieved from culture supernatant. The specific activity and recovery yield after purification were 294.0 U/mg and 40.15%, respectively, which were comparable with other contemporary protocols. The homogeneity of the enzyme was tested through sodium dodecyl sulphate polyacrylamide gel electrophoresis and a single band of 56.3 kDa was observed. Zymogram analysis finally confirmed the occurrence of endoglucanase in the single band.

 

 

Indian Journal of Experimental Biology

Vol. 51, November 2013, pp. 960-968

 

 

Analysis of alteration of gut microbial population under the exposure of graded hyperbaric pressures: Application of metagenomic approach

Chiranjit Maity, Atanu Adak, Suman Kumar Halder, Arijit Jana, Kuntal Ghosh, Pradeep Kumar Das Mohapatra,
Bikas Ranjan Pati & Keshab Chandra Mondal*

Department of Microbiology, Vidyasagar University, Midnapore 721 102, India

Received 8 December 2012; revised 18 February 2013

Gastroenterological disorders are very common at hyperbaric conditions. The present study was conducted to find out the impact of gut flora on the gastrointestinal disorders created at such environmental circumstances. For this, male albino rat were exposed to graded hyperbaric pressures (915 and 1277 mmHg) and large intestinal content was examined for microbial composition using culture based and PCR-DGGE tools. After 30 day exposure, total aerobes (38.54 and 375.57 folds, 1.35 and 1.58 gdi) and E. coli (126.05 and 873.23 folds, 1.31 and 1.44 gdi) were increased whereas total anaerobes (7.01 × 104 and 8.84 × 103 folds, -1.56 and -1.39 gdi), Enterobacter spp. (-2.45 and -1.00 gdi) and Clostridium perfringens (12.88 and 54.16 folds, -1.38 and -1.75 gdi) were decreased significantly in respect to control after exposure of simulated hyperbaric pressures like at 915 and 1277 mmHg, respectively. Metagenomics study revealed an overall reduction in total microbial profile was noted than control at higher level hyperbaric pressure, i.e., 1277 mmHg air pressure for highest duration of exposure. Though, some new bands also appeared which indicated the expansion of dormant or new microbiota, Variation in the numbers of these newly dominated bacteria was correlated to dose and duration of hyperbaric treatment. The histological results clearly indicated that hyperbaric environment induced severe inflammation in the mucosal and submucosal layer of large intestine. Thus, the result suggest that hyperbaric pressure is an important exogenous factor that strongly modulated the intestinal morphology and microbial ecology, and induced several gastrointestinal ailments during hyperbarism.

 

 

 

Indian Journal of Experimental Biology

Vol. 51, November 2013, pp. 969-972

 

 

Biosynthesis of colloidal gold nanoparticles by Streptomyces sp. NK52 and
its anti-lipid peroxidation activity

Divya Prakash1, Vishal Mahale2, Ashok Bankar2, Neelu Nawani3*, Smita Zinjarde2 & Balasaheb Kapadnis1

1Department of Microbiology, University of Pune, Pune 411007, India

2Institute of Bioinformatics and Biotechnology, University of Pune, Pune 411007, India

3Dr. D. Y. Patil Biotechnology and Bioinformatics Institute, Dr. D. Y. Patil Vidyapeeth, Pune 411 033, India

Received 8 December 2012; revised 18 February 2013

Gold nanoparticles (Au-NPs) were synthesized from chloroauric acid using cell free supernatant of Streptomyces sp. NK52 grown in nutrient broth. These nanoparticles were synthesized by varying pH and temperature of the reaction mixture and chloroauric acid concentration. The nanoparticles were characterized by spectrometry, X-ray diffraction, Scanning electron microscopy and energy dispersive spectrometry. Au-NP ranged from 10-100 nm in size and exhibited a polydispersive nature with various shapes like rods, hexagons, triangles, spheres. The diffraction peaks at 2θ = 38.1◦ and 44.5◦ could be assigned to the (1 1 1) and (2 0 0) planes of a faced centre cubic (fcc) lattice of gold. Au-NP showed 47% inhibition of lipid peroxidation in vitro. To the best of our knowledge, this is the first report on the rapid biosynthesis of Au-NP using cell free supernatant of Streptomyces sp. and its evaluation for anti-lipid peroxidation.

 

 

Indian Journal of Experimental Biology

Vol. 51, November 2013, pp. 973-978

 

 

Biosynthesis and characterization of mercury sulphide nanoparticles
produced by Bacillus cereus MRS-1

 

Sundararaju Sathyavathi, Arumugam Manjula, Jeyaprakash Rajendhran  & Paramasamy Gunasekaran*

Department of Genetics, School of Biological Sciences, Madurai Kamaraj University, Madurai 625021, India

Received 9 December 2012; revised 18 February 2013

Mercury is a highly toxic heavy metal accumulated in the environment, which can be detoxified by reducing Hg2+ to non toxic form. Bacteria resistant to toxic metals and capable of converting them into non toxic forms have a direct application in the bioremediation of contaminated sites. In this study, mercury resistant strain Bacillus cereus MRS-1 was isolated from electroplating industrial effluent. This strain exhibited the ability to convert mercury into extracellular sulphide nanoparticles of mercury. The recovered HgS nanoparticles have been characterized by UV-VIS spectrophotometer, FT-IR, atomic force microscopy, transmission electron microscopy, scanning electron microscopy, energy dispersive X-ray analysis, powder X- ray diffraction pattern and thermogravimetric analysis. The synthesized nanoparticles were spherical with a size range of 10–100 nm. This strain can be potentially exploited for the production of HgS nanoparticles as well as for detoxification of mercury in the environment without producing secondary pollution of mercury methylation or Hg (0) volatilization.

 

 

Indian Journal of Experimental Biology

Vol. 51, November 2013, pp. 979-983

 

Optimization of lipid enriched biomass production from oleaginous fungus using response surface methodology

S P Jeevan Kumar & Rintu Banerjee*

Microbial and Downstream Processing Laboratory

Department of Agricultural and Food Engineering

Indian Institute of Technology Kharagpur 721 302, India

Received 9 December 2012; revised 22 February 2013

Oleaginous microorganisms have emerged as potential sources of oils for biodiesel production. To replenish as an alternative to the vegetable oils, higher lipid accumulating strain coupled with process optimization is indispensable. In the present study, response surface methodology (RSM) based central composite design (CCD) was used for optimization of lipid content from oleaginous fungus Aspergillus sp. Maximum lipid yield of 73.07% (w/w) was achieved at 3% (v/v) inoculum volume, pH 5, glucose 1% (w/v), urea 0.5 % (w/v) and incubation time of 5 (days). Biomass (2.08 g/L) having a lipid content of 73.07 % (w/w) with major constituents of hexadecanoic acid methyl ester and 9-Octadecenoic acid methyl ester were obtained. The lipid composition signifies that from the oleaginous microbe are highly encouraging and desirable to be considered as diesel substitute.

 

 

Indian Journal of Experimental Biology

Vol. 51, November 2013, pp. 984-991

 

 

Characterization and antimicrobial activity of lectins from Penicillium sp.

R S Singh*, P Jain & H P Kaur

Carbohydrate and Protein Biotechnology Laboratory, Department of Biotechnology,
Punjabi University, Patiala 147 002, India

Received 9 December 2012; revised 18 February 2013

Ten Penicillium sp. were screened for lectin activity for occurrence of lectins. Mycelial extracts from submerged cultures of P. corylophilum, P. expansum and P. purpurogenum showed agglutination against human (A, B, AB and O), goat, sheep, pig and rabbit erythrocytes. Neuraminidase treatment to human blood type O erythrocytes substantially increased their agglutinability by all the lectins as compared to untreated erythrocytes. Modification of erythrocyte surfaces by protease increased the lectin titre only of P. corylophilum with no effect on other two lectins. P. corylophilum and P. expansum displayed relatively lower titres in mycelial extracts prepared from agar plate cultures as compared to broth cultures. A panel of sugars was tested for inhibition of lectin activity. All the lectins were found to be specific for asialofetuin, bovine submaxillary mucin, porcine stomach mucin, chondroitin-6-sulphate, D-sucrose and D-glucose. P. corylophilum lectin was expressed (Titre 8) by 5 day old cultures, reaching its maximum level (Titre 32) upon 8 days of cultivation, thereafter declin in lectin activity was observed. P. purpurogenum lectin was expressed by 7-10 days old cultures, while in P. expansum maximum lectin activity was elaborated by 5-8 days old cultures. Lectin extracts from all the three species were found to possess antimicrobial activities. Lectin extracts from the three Penicillium species displayed antifungal activity and antibacterial activity against Gram-negative and Gram-positive bacterial strains.

 

 

Indian Journal of Experimental Biology

Vol. 51, November 2013, pp. 992-996

 

 

Optimization of enzymatic saccharification of microwave pretreated sugarcane tops through response surface methodology for biofuel

Devendra Prasad Maurya, Siddharth Vats , Sudheer Rai & Sangeeta Negi*

Motilal Nehru National Institute of Technology, Allahabad 211 004, India

Received 9 December 2012; revised 18 February 2013

The optimization of biomass loading enzyme loading, surfactant concentration and incubation time, using response surface methodology (RSM) and Box Behnken design for enzymatic saccharification of sugarcane tops (SCT) for maximum recovery of fermentable sugars using crude cellulases, resulted in 90.24% saccharification efficiency. Maximum saccharification yield of 0.376 g/g glucose as substrate for ethanol production was observed at optimal conditions of 10% biomass loading (pretreated), 100FPU/g of cellulase loading, 0.04% (w/w) surfactant concentration and 72 h of incubation time.

 

 

Indian Journal of Experimental Biology

Vol. 51, November 2013, pp. 997-1003

 

 

A comparative study on cell disruption methods for release of aspartase
from E. coli K-12

R S Singh

Carbohydrate and Protein Biotechnology Laboratory, Department of Biotechnology,
Punjabi University, Patiala 147 002, India

Received 10 December 2012; revised 2 March 2013

Applicability of different mechanical cell disruption techniques namely sonication, bead milling and French press for the release of aspartase from E. coli K-12 was compared. Various operating parameters of each technique were optimized to obtain maximum aspartase release. The efficiency of aspartase release and cell disruption by all the methods was also compared under optimal conditions. The maximum release of aspartase (98.22%) and maximum cell breakage (84.25%) was observed using French press, while 92% of aspartase release was obtained by both sonication and bead milling. The order of cell disruption constant (k) for aspartase release by these methods was French press > bead milling > sonication. Disruption of cells using French press also demonstrated maximum protein release (14.12 mg/mL). The crude enzyme preparations can be further used for purification and its applications.

 

 

Indian Journal of Experimental Biology

Vol. 51, November 2013, pp. 1004-1011

 

 

Isolation of a lead tolerant novel bacterial species, Achromobacter sp. TL-3: Assessment of bioflocculant activity

Neha Batta1, Sanjukta Subudhi1*, Banwari Lal1* & Arundhuti Devi2

1Environmental and Industrial Biotechnology Division, The Energy and Resources Institute (TERI), Habitat Place,
Darbari Seth Block, Lodhi Road, New Delhi 110 003, India

2Institute of Advanced Study in Science and Technology, Vigyan Path, Paschim Boragaon, Guwahati 781 035, India

Received 6 December 2012; revised 2 March 2013

Lead is one of the four heavy metals that has a profound damaging effects on human health. In the recent past there has been an increasing global concern for development of sustainable bioremediation technologies for detoxification of lead contaminant. Present investigation highlights for lead biosorption by a newly isolated novel bacterial species; Achromobacter sp. TL-3 strain, isolated from activated sludge samples contaminated with heavy metals (collected from oil refinery, Assam, North-East India). For isolation of lead tolerant bacteria, sludge samples were enriched into Luria Broth medium supplemented separately with a range of lead nitrate; 250, 500, 750, 1000, 1250 and 1500 ppm respectively. The bacterial consortium that could tolerate 1500 ppm of lead nitrate was selected further for purification of lead tolerant bacterial isolates. Purified lead tolerant bacterial isolates were then eventually inoculated into production medium supplemented with ethanol and glycerol as carbon and energy source to investigate for bioflocculant production. Bioflocculant production was estimated by monitoring the potential of lead tolerant bacterial isolate to flocculate Kaolin clay in presence of 1% CaCl2. Compared to other isolates, TL-3 isolate demonstrated for maximum bioflocculant activity of 95% and thus was identified based on 16S rRNA gene sequence analysis. TL3 isolate revealed maximum homology (98%) with Achromobacter sp. and thus designated as Achromobacter sp. TL-3. Bioflocculant activity of TL-3 isolate was correlated with the change in pH and growth. Achromobacter sp. TL-3 has significant potential for lead biosorption and can be effectively employed for detoxification of lead contaminated waste effluents/waste waters.

 

 

Indian Journal of Experimental Biology

Vol. 51, November 2013, pp. 1012-1017

 

 

Evaluation of polymeric adsorbent resins for efficient detoxification of liquor generated during acid pretreatment of lignocellulosic biomass

 

Soolamkandath Variem Sandhya, Kumar Kiran, Mathiyazhakan Kuttiraja, Varghese Elizabeth Preeti,
Raveendran Sindhu, Sankar Vani, Sukumaran Rajeev Kumar, Ashok Pandey
& Parameswaran Binod*

Centre for Biofuels, CSIR-National Institute for Interdisciplinary Science and Technology (NIIST),
Trivandrum 695 019, India

Received 9 December 2012; revised 9 February 2013

Production of fuel ethanol from lignocellulosic biomass conventionally includes biomass pretreatment, hydrolysis, and fermentation. The liquor generated during dilute acid pretreatment of biomass contains considerable quantities of pentose sugars as well as various degradation products of sugars and lignin, like furfural, hydroxymethyl furfural (HMF), organic acids, aldehydes and others, which are known to be inhibitory for microbial growth. This pentose rich liquor is a potent resource which can be used to produce alcohol or other value added metabolites by microbial fermentation. However, the presence of these inhibitory compounds is a major hindrance and their removal is essential for efficient utilization of this byproduct stream. In the present work, the polymeric adsorbent resins, XAD-4, XAD-7 and XAD-16 were evaluated for their ability to adsorb fermentation inhibitors like furfural and HMF from the acid pretreated liquor. These resins could remove 55-75% of furfural and 100% of HMF and more than 90% sugar remained un-adsorbed in the pretreated liquor. Desorption of furfural from stationary phase was evaluated by using ethanol and hot water. The results suggest that these polymeric resins may be used for detoxification of acid pretreatment liquor with selective removal of sugar degradation products without affecting the sugar content in the solution.

 

 

Indian Journal of Experimental Biology

Vol. 51, November 2013, pp. 1018-1023

 

 

Treatment of waste gas containing low concentration of dimethyl sulphide
in a high performance biotrickling filter

Balendu Shekher Giri, Asha Juwarkar, S N Mudliar & R A Pandey*

CSIR-National Environmental Engineering Research Institute, (CSIR-NEERI), Nehru Marg, Nagpur 440 020, India

Received 9 December 2012; revised 11 February 2013

A bench-scale biotrickling filter was operated in the laboratory for the treatment of dimethyl sulphide (DMS).
The biotrickling filter was packed with pre-sterilized polyurethane foam and seeded with biomass developed from garden soil enriched with DMS. The biotrickling filter was operated for the generation of process parameters. The biotrickling filter could remove an average removal efficiency of 40.95 % at an effective bed contact time of 84 sec with an average loading rate of 0.56 mg/m3/h. Evaluation of microbiological status of the biotrickling filter indicated the presence of other bacterial cultures viz. Paenibacillus polymyxa, and Bacillus megaterium, besides Bacillus sphaericus.

 

 

Indian Journal of Experimental Biology

Vol. 51, November 2013, pp. 1024-1031

 

 

Proteolytic enzyme mediated antagonistic potential of Pseudomonas aeruginosa against Macrophomina phaseolina

 

Devaraj Illakkiam, Nishanth Lipton Anuj, Paramasivan Ponraj, Manoharan Shankar,
Jeyaprakash Rajendhran & Paramasamy Gunasekaran*

Department of Genetics, Centre for Excellence in Genomic Sciences, School of Biological Sciences,
Madurai Kamaraj University, Madurai 625 021, India

Received 7 December 2012; revised 18 February 2013

A new antagonistic bacterial strain PGPR2 was isolated from the mungbean rhizosphere and documented for the production of hydrolytic enzymes with antifungal activity. Based on the phylogenetic analysis of the 16S rRNA gene sequence and phenotyping, this strain was identified as Pseudomonas aeruginosa. Maximum protease activity (235 U/mL) was obtained at 24 h of fermentation. The protease was purified to homogeneity in three steps: ammonium sulphate precipitation, anion exchange chromatography on DEAE- cellulose resin and gel filtration chromatography using P6 column. The purified enzyme had a molecular weight of ~33 kDa. The purified protease exhibited maximum activity at pH 6.0 and retained 80% of activity in a pH range of 5.0 - 9.0. Proteolytic activity was maximum in a temperature range of 40–70 °C. However, the enzyme was stable at 40 °C for 60 min. Among the metals tested, Mg2+ enhanced the protease activity. Internal amino acid sequence of the protease obtained by MALDI -ToF and subsequent Mascot database search showed maximum similarity to the HtpX protease of P. aeruginosa strain PA7. Thus, by virtue of its early production time, thermostability and effective antifungal ability, the protease purified and characterized from P. aeruginosa PGPR2 has several potential applications as fungicidal agents in agriculture.

 

 

Indian Journal of Experimental Biology

Vol. 51, November 2013, pp. 1032-1037

 

 

Candida albicans biofilm inhibition by synergistic action of terpenes
and fluconazole

Suma C Pemmaraju, Parul A Pruthi, R Prasad & Vikas Pruthi*

Department of Biotechnology, Indian Institute of Technology Roorkee, Roorkee 247 667, India

Received 8 December 2012; revised 18 February 2013

The current treatment options for Candida albicans biofilm-device related infections are very scarce due to their intrinsic increased tolerance to antimycotics. The aim of this work was to study synergistic action of terpenes (eugenol, menthol and thymol) with fluconazole (FLA) on C. albicans biofilm inhibition. The minimum inhibitory concentration (MIC) assayed using CLSI M27-A3 broth micro-dilution method showed antifungal activity against C. albicans MTCC 227 at a concentration of 0.12 % (v/v) for both thymol and eugenol as compared to 0.25 % (v/v) for menthol. FLA was taken as positive control. The effect of these terpenes on metabolic activity of preformed C. albicans biofilm cells was evaluated using 2,3-bis (2-methoxy-4-nitro-5-sulfophenyl)-2H-tetrazolium-5-carboxanilide (XTT) reduction assay in 96-well polystyrene microtiter plate. Thymol and eugenol were more effective at lower concentrations of ≥ 1.0 % (v/v) than menthol. Synergistic studies using checkerboard micro-dilution assay showed fractional inhibitory concentration index
(Σ FIC=0.31) between thymol/FLA followed by eugenol/FLA (Σ FIC=0.37) and menthol/FLA (Σ FIC<0.5)
against
pre-formed C. albicans biofilms. Thymol with fluconazole showed highest synergy in reduction of biofilm formation than eugenol and menthol which was not observed when their activities were observed independently. Adherence assay
showed 30% viability of C. albicans cells after 2 h of treatment with 0.05 % (v/v) thymol/FLA. Effect of thymol/FLA on C. albicans adhesion visualized by SEM micrographs showed disruption in number of candidal cells and alteration in structural design of C. albicans. Thus, the study demonstrated synergistic effect of terpenes with fluconazole on C. albicans biofilm, which could be future medications for biofilm infections.

 

 

Indian Journal of Experimental Biology

Vol. 51, November 2013, pp. 1038-1045

 

 

Liquid based formulations of bacteriophages for the management of waterborne bacterial pathogens in water microcosms

Sangeeta Ahiwale1, Sujata Tagunde2, Sushama Khopkar3, Mrudula Karni3, Milind Gajbhiye2 & Balasaheb Kapadnis2 *

1Department of Environmental Sciences, University of Pune, Pune 411 007, India.
2Department of Microbiology, University of Pune, Pune 411 007, India

3Department of Microbiology, M. P. College, Pimpri, Pune 400 017

Received 9 December 2012; revised 2 March 2013

Water resources are contaminated by life-threatening multidrug resistant pathogenic bacteria. Unfortunately, these pathogenic bacteria do not respond to the traditional water purification methods. Therefore, there is a need of environmentally friendly strategies to overcome the problems associated with the antimicrobial resistant bacterial pathogens. In the present study, highly potent lytic phages against multidrug-resistant Salmonella enterica serovar Paratyphi B, Pseudomonas aeruginosa and Klebsiella pneumoniae were isolated from the Pavana river water. They belonged to the Podoviridae and Siphoviridae families. These phages were purified and enriched in the laboratory. Monovalent formulations of φSPB, BVPaP-3 and KPP phages were prepared in three different liquids viz., phage broth, saline and distilled water. The phages were stable for almost 8-10 months in the phage broth at 4 °C. The stability of the phages in saline and distilled water was 5-6 months at 4 °C. All of the phages were stable only for 4-6 months in the phage broth at 30 °C. The monovalent phage formulation of φSPB was applied at MOI < 1, as disinfectant against an exponential and stationary phase cells of Salmonella enterica serovar Paratyphi B in various water microcosms. The results indicated that there was almost 80 % reduction in the log phase cells of Salmonella serovar Paratyphi B in 24 h. In stationary phase cells, the reduction was comparatively less within same period. At the same time, there was concomitant increase in the phage population by 80% in all the microcosms indicating that φSPB phage is highly potent in killing pathogen in water. Results strongly support that the formulation of φSPB in the phage broth in monovalent form could be used as an effective biological disinfectant for preventing transmission of water- borne bacterial pathogens, including antimicrobial resistant ones.