Indian Journal of Biotechnology

 

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VOLUME 9
CODEN: IJBNAR 9(1) (2010) 1-110

NUMBER 1

JANUARY 2010
ISSN: 0972-5849 (print),
0975-0967 (online)

 

 

   
CONTENTS
 
   

Reviews

 

RAPD marker system in insect study: A review

7

        Subodh Kumar Jain, Bharat Neekhra, Divya Pandey & Kalpana Jain

 

 

 

Proteomic approach to autoimmune disorders: A review

13

        Vandana D Pradhan, Neha R Deshpande & K Ghosh

 

 

 

Papers

 

Borrelidin: A prospective drug

18

        D V R N Bhikshapathi, Y Shravan Kumar, Y Madhusudan Rao & V Kishan

 

 

 

Comparative analysis of transcription factors of insulin signaling (Insulin receptor substrate family)

24

        Sampoornam Balakrishnan, Satyavani Kumpatla & Vijay Viswanathan

 

 

 

cDNA cloning, sequencing, and expression of a- and b-neurotoxins from Thai-Malayan krait

31

        Sunutcha Suntrarachun, Thaweesak Tirawatnapong, Suchittra Khunsap &
Sunanta Puempumpanich

 

 

 

Strain differentiation and characterization of canine parvovirus by PCR and RE mapping

38

        S Nandi, R Anbuzhagan & Manoj Kumar

 

 

 

Assessment of genetic diversity in Indian Perilla [Perilla frutescens (L.) Britton] landraces
using STMS markers

43

        Nidhi Verma, M K Rana, K S Negi, Gunjeet Kumar, K V Bhat, Y J Park & I S Bisht

 

 

 

Evaluation of genetic diversity in Jatropha curcas L. using RAPD markers

50

        Ikbal, K S Boora & R S Dhillon

 

 

 

Identification of RAPD markers linked to plant type gene in pigeonpea

58

        P Dhanasekar, K N Dhumal & K S Reddy

 

 

 

Investigation on relative genome sizes and ploidy levels of Darjeeling-Himalayan
Rhododendron species using flow cytometer

64

        Kalyan Kumar De, Aniruddha Saha, Ranju Tamang & Bilok Sharma

 

 

 

Evaluation and optimization of DNA extraction method for Dalbergia sissoo leaf

69

        H S Ginwal & Shalini Singh Maurya

 

 

 

Differentiation of petroleum hydrocarbon-degrading Pseudomonas spp. based on
lectin binding of cell extracts in an agglutination assay

74

        Bhaben Tanti & Alak Kumar Buragohain

 

   

Antibiotic resistance profile of halophilic microorganisms isolated from tannery effluent

80

        Rakesh Ghosh, Pijush Kanti Chattopadhyay, Budhhadeb Chattopadhyay & Debasish Pal

 

 

 

Production of fibrolytic enzymes in repeat-batch culture using immobilized zoospores of
anaerobic rumen fungi

87

        Manpal Sridhar & Deepak Kumar

 

 

 

Homology modeling and docking studies between HIV-1 protease and carbamic acid

96

        M Balakrishnan, R C Srivastava & Mayank Pokhriyal

 

 

 

Regeneration of multiple shoots from the callus cultures of Macrotyloma uniflorum (Lam.) Verdc.

101

        D H Tejavathi, V R Devaraj, Savitha M Murthy, P Anitha & R Nijagunaiah

 

 

 

Instructions to Contributors

107

 

 

          AUTHOR INDEX

 

 


Anbuzhagan R

38

Anitha P

101

 

 

Balakrishnan M

96

Balakrishnan S

24

Bhat K V

43

Bhikshapathi D V R N

18

Bisht I S

43

Boora K S

50

Buragohain A K

74

 

 

Chattopadhyay B

80

Chattopadhyay P K

80

 

 

De K K

64

Deshpande N R

13

Devaraj V R

101

Dhanasekar P

58

Dhillon R S

50

Dhumal K N

58

 

 

Ghosh K

13

Ghosh R

80

Ginwal H S

69

Ikbal

50

 

 

Jain K

7

Jain S K

7

 

 

Khunsap S

31

Kishan V

18

Kumar D

87

Kumar G

43

Kumar M

38

Kumar Y S

18

Kumpatla S

24

 

 

Maurya S S

69

Murthy S M

101

 

 

Nandi S

38

Neekhra B

7

Negi K S

43

Nijagunaiah R

101

 

 

Pal D

80

Pandey D

7

Park Y J

43

Pokhriyal M

96

Pradhan V D

13

Puempumpanich S

31

 

 

Rana M K

43

Rao Y M

18

Reddy K S

58

 

 

Saha A

64

Sharma B

64

Sridhar M

87

Srivastava R C

96

Suntrarachun S

31

 

 

Tamang R

64

Tanti B

74

Tejavathi D H

101

Tirawatnapong T

31

 

 

Verma N

43

Viswanathan V

24


 

 

 

Indian Journal of Biotechnology

Vol 9, January 2010, pp 7-12

 

RAPD marker system in insect study: A review

Subodh Kumar Jain*, Bharat Neekhra, Divya Pandey and Kalpana Jain

Department of Biotechnology, Dr H S Gour University, Sagar 470 003, India

Received 30 September 2008; revised 2 June 2009; accepted 7 August 2009

Insects represent a major life form on earth. So far, nearly 0.9 million insect species are discovered, comprising 75% of all the recorded animal species. Some of the insect species are easy to identify and categorize, while for others, it is difficult because of their small size and morphological similarity. Moreover, it is further difficult to identify morphological variation due to environmental factors by available traditional methods. To overcome these problems, the advanced molecular techniques, viz., PCR (Polymerase Chain Reaction), RFLP (Restriction Fragment Length Polymorphism), RAPD (Random Amplified Polymorphic DNA), and AFLP (Arbitrary Fragment Length Polymorphism) have been a great help. RAPD markers have been used in gene mapping to characterize cultivars and species genetically, infer phylogeny and biogeography of insect population and understand modes of evolution and evolutionary trajectories. Thus, RAPD markers have become the most common yardsticks for measuring genetic differences between individuals, within and between related species or population. The unprecedented advancements in modern molecular biology, particularly in those of DNA marker technology, have created a wealth of technical know-how that finds useful application in molecular ecology research in insects.

Keywords: Genetic variation, insects, molecular markers, PCR, RAPD

 

 

Indian Journal of Biotechnology

Vol 9, January 2010, pp 13-17

 

Proteomic approach to autoimmune disorders: A review

Vandana D Pradhan, Neha R Deshpande and K Ghosh

Department of Autoimmune Disorders, National Institute of Immunohaematology, King Edward Memorial Hospital
Parel, Mumbai 400 012, India

Received 23 February 2009; revised 25 May 2009; accepted 28 July 2009

Proteomics is the study of structural and functional endowment of cells, tissues or organs. This science brings together powerful tools—physical separation techniques like 2-D electrophoresis and mass spectroscopy. It also includes various monoclonal antibodies and other probes coupled with which analysis is done by systems biology approach using modern software. Various statistical, probabilistic, humanistic and artificial neural network algorithms and at the same time incorporating elements of chaos and fractal theories are used to study the interactions of multitude of proteins in the cell. This allows separation of large background noise of high concentration proteins inside the cell from pathobiologically and aetiologically relevant protein molecules present in nano, femto or even atto molar concentrations. Pattern recognition algorithms in modern proteomic techniques will help in understanding aetiopathogenesis of disease, discovering diagnostically and prognostically important biomarkers and molecular targets for future drug discovery. These techniques will have important applications in autoimmune disorders and other disorders which are in general difficult to manage. The present review shows how proteomic approach can illuminate various facets of these groups of elusive disorders
in near future.

Keywords: Proteomics, autoimmune disorders, autoantibodies, biomarkers, bioinformatics

 

 

Indian Journal of Biotechnology

Vol 9, January 2010, pp 18-23

Borrelidin: A prospective drug

D V R N Bhikshapathi, Y Shravan Kumar, Y Madhusudan Rao and V Kishan*

University College of Pharmaceutical Sciences, Kakatiya University, Warangal 506 009, India

Received 23 December 2008; revised 20 April 2009; accepted 25 June 2009

Borrelidin, an antibiotic isolated from Streptomyces species and firstly isolated from Streptomyces rochei, is a possible drug candidate due to its recently reported antiangiogenic activity and other biological activities. Keeping in view the potential scope of the antibiotic, we performed computational studies like application of Lipinski’s rule for borrelidin, and found from the properties of the borrelidin that it possessed good absorption or permeability across the biological membrane. Influence of borrelidin on CYP3A enzyme was tested by pretreatment of the male Wistar rats at a concentration of 100 µg/mL/rat daily for 5 d by oral administration, buspirone was used as a substrate; the results indicated the role of borrelidin acting as an inhibitor of CYP3A. The effect of borrelidin on pretreated rat liver by micromorphological studies was also done and results showed the damage on liver with borrelidin pretreatment. Influence of borrelidin on platelets was studied at three different concentrations along with known antiplatelet aggregating agent, aspirin. Platelet aggregation was noticed for all the three tested concentrations in comparison to control. Influence of borrelidin on prothrombin time was also performed and compared with known anticoagulant, warfarin sodium, have little effect of borrelidin on the prothrombin time which indicated the anticoagulant activity.

Keywords: Borrelidin, Lipinski’s rule, CYP3A enzyme, platelets, prothrombin time

 

 

Indian Journal of Biotechnology

Vol 9, January 2010, pp 24-30

 

Comparative analysis of transcription factors of insulin signaling (Insulin receptor substrate family)

Sampoornam Balakrishnan, Satyavani Kumpatla and Vijay Viswanathan*

M V Hospital for Diabetes and Diabetes Research Centre, Chennai, 600 013 India

Received 11 November 2008; revised 23 March 2009; accepted 8 June 2009

The aim of this study was to identify the role of insulin receptor substrates (IRS) in insulin signaling by doing sequence based comparative analysis. The sequences of IRS family (1, 2, 4, 5 & 6) were submitted to predict protein motif scan and clustal W for motif, domain identification and multiple sequence alignment. All the 5 IRS had some common motifs like amidation, Asn glycosylation, myristoylation, cAMP, casein kinase II, protein kinase C phosphorylation sites and functional domains like PTB and PH. Some functional motifs like tyrosine phosphorylation, Rho kinase α binding, OGFR, PSGP were identified in IRS-1, RGD motifs in IRS-1 and 4, XYPPX motif and ECM protein domain in IRS-2, NLS motif and extension like domain in IRS-2 and 4, ascorbate cytosolic peroxidase, class II peroxidase superfamily domain, NHL and 2 octapeptide repeats and ATP_GTP_ attachment sites in IRS-4. These domains and motifs may have inducing and inhibitory effects on insulin signaling.

Keywords: signaling pathway, insulin signaling, IRS, Rho kinase a, upregulation, downstream signaling

 

 

Indian Journal of Biotechnology

Vol 9, January 2010, pp 31-37

 

cDNA cloning, sequencing, and expression of a- and b-neurotoxins from
Thai-Malayan krait

Sunutcha Suntrarachun*, Thaweesak Tirawatnapong1, Suchittra Khunsap and Sunanta Puempumpanich

Research and Development, Queen Saovabha Memorial Institute, The Thai Red Cross Society, Patumwan, Bangkok 10330, Thailand

1Faculty of Medicine, Chulalongkorn University, Patumwan, Bangkok 10330, Thailand

Received 29 May 2008; revised 25 march 2009, accepted 19 June 2009

Amplification products of a- and b-neurotoxin from Thai-Malayan krait (Bungarus candidus) were cloned and expressed in TA expression vector. The expressed protein could not be distinguished by SDS-PAGE. Hence, immunoblotting was performed using AntiHis (C-term)-HRP antibody. The antibody could identify the histidine tags at 24 h incubation with 0.02% L-arabinose. To increase the expression level, PCR products were cloned into PCR2.1 cloning vector and pGEX2T expression vector. The optimal condition for protein expression was IPTG induction at 1 mM for 24 h. Neurotoxin fusion proteins were used as antigen to generate antibodies in mice. In vitro neutralization indicated that antibody against neurotoxin fusion proteins raised in mice was able to neutralize 2´ LD50 of crude venom. This result provides basic data for the use of the neurotoxin fusion proteins as immunogens in the development of specific antivenoms against the B. candidus venom.

Keywords: expression, induction, neurotoxin fusion proteins, recombinant proteins

 

 

Indian Journal of Biotechnology

Vol 9, January 2010, pp 38-42

 

Strain differentiation and characterization of canine parvovirus by PCR and RE mapping

S Nandi*, R Anbuzhagan and Manoj Kumar

Centre for Animal Disease Research and Diagnosis (CADRAD)
Indian Veterinary Research Institute (IVRI), Izatnagar 243 122, India

Received 20 January 2009; revised 1 June 2009; accepted 6 August 2009

Canine parvovirus 2 (CPV2) is a highly contagious and fatal disease of dogs, causing acute hemorrhagic enteritis and myocarditis. In this study, different mutant of CPV2 prevalent in India have been isolated in Madin-Darby canine kidney (MDCK) cells and characterized by polymerase chain reaction (PCR) and restriction endonuclease (RE) mapping. The faecal samples from gastroenteritic dog suspected for CPV2 infection were collected in a suitable medium, processed and inoculated in MDCK cells. The viral DNA from faecal samples was extracted using phenol-chloroform method. PCR were carried out with two different primer pairs, pCPV-2ab and pCPV-2b, to distinguish the strain prevalent in field condition. The primer pCPV-2ab recognized both the variant CPV-2a and CPV-2b, whereas the primer pCPV-2b recognized only the variant CPV-2b, enabling the differentiation of CPV-2a variant from CPV-2b in field isolates. The different PCR products were further analyzed by using RE mapping.

Keywords: Canine parvovirus, haemorrhagic enteritis, MDCK, PCR, RE mapping

 

 

Indian Journal of Biotechnology

Vol 9, January 2010, pp 43-49

 

Assessment of genetic diversity in Indian Perilla [Perilla frutescens (L.)
Britton] landraces using STMS markers

Nidhi Verma1, M K Rana1, K S Negi2, Gunjeet Kumar1, K V Bhat1, Y J Park3 and I S Bisht1*

1National Bureau of Plant Genetic Resources, New Delhi 110 012, India

2National Bureau of Plant Genetic Resources, Regional Station, Bhowali 263 132, India

3National Institute of Agricultural Biotechnology, RDA, Suwon, 441-744, Republic of Korea

Received 21 October 2008; revised 15 April 2009; accepted 30 June 2009

Inter-population diversity in 54 Indian Perilla landraces and 18 accessions of exotic origin was investigated using sequence tagged microsatallite (STMS) markers. The STMS markers clearly distinguished the Indian accessions from the exotic ones. Neighborhood-joining (NJ) clustering pattern revealed association of geographical diversity and genetic diversity for Indian Perilla germplasm. Population genetic parameters studied for 14 Indian Perilla populations from two distinct regions (North-eastern region and North-western Himalayas, Uttarakhand State) revealed greater diversity for accessions from the later region. Analysis of molecular variance, revealed that bulk of the variations existed between populations within groups, followed by variations within populations and between groups. The summary of group-wise F-statistics and gene flow revealed greater population differentiation for Uttarakhand populations as compared to accessions from North-eastern region of India.

Keywords: Perilla frutescens, genetic diversity, Indian Himalayas, molecular characterization, STMS markers

 

 

Indian Journal of Biotechnology

Vol 9, January 2010, pp 50-57

 

Evaluation of genetic diversity in Jatropha curcas L. using RAPD markers

Ikbal1, K S Boora1 and R S Dhillon2*

1Department of Biotechnology and Molecular Biology, and 2Department of Forestry
CCS Haryana Agricultural University, Hisar 125 004, India

Received 3 October 2008; revised 24 March 2009; accepted 12 June 2009

Random amplified polymorphic DNA markers were used to evaluate the genetic diversity in a representative population of Jatropha curcas L. from different eco-geographical regions of India. Out of 50 dacamer primers used, 44 yielded polymorphic banding pattern. A total of 328 DNA bands were obtained, of which 308 (93.90%) were polymorphic. The polymorphism was scored and used in band sharing analysis to identify genetic relationship. Cluster analysis based on Jaccard’s similarity coefficient using UPGMA grouped all the 40 genotypes into two major groups at a similarity coefficient of 0.54. Similarity indices ranged from 0.44 to 0.92 with an average of 0.73, indicating a moderate to high genetic variability among the genotypes. The highest similarity coefficient was detected between accessions JC-32 and JC-35 from Madhya Pradesh, and the lowest in accessions JC-9, JC-14 and JC-19 from Rajasthan (Pipalkhunt, Banswara), Punjab (Kandi area) and Haryana (Ladwa, Hisar), respectively. Also, the Jatropha populations from diverse agro-climatic regions were more dispersed on the principal coordinates plot, revealing a wide genetic base.

Keywords: Genetic variation, Jatropha curcas, RAPD

 

 

Indian Journal of Biotechnology

Vol 9, January 2010, pp 58-63

 

Identification of RAPD markers linked to plant type gene in pigeonpea

P Dhanasekar1*, K N Dhumal2 and K S Reddy1

1Nuclear Agriculture and Biotechnology Division, Bhabha Atomic Research Centre, Trombay, Mumbai 400 085, India

2Department of Botany, University of Pune, Pune 411 007, India

Received 3 December 2008; revised 20 May 2009; accepted 27 July 2009

Pigeonpea is the second most important pulse crop of India accounting for almost 90% of the World’s pigeonpea area and production. Altering the plant types could improve the economic yield and its adaptability as well. Molecular markers can identify and tag plant type genes without much environmental interference. In the present investigation of identifying and tagging RAPD markers linked with the plant type trait, the plant material consisted of parents, F1 and a F2 population obtained by crossing genotypes TT44-4 with open-tall and TDI2004-1 with compact-dwarf plant types. RAPD analysis was performed on 84 genotyped F2 plants following a bulked segregant analysis (BSA) approach. In the parental screening with 200 random decamer primers, only eight showed polymorphism, of which only two could be linked to the plant type gene. Two markers (OPF04700 and OPA091375) were identified that were present in the open-tall plants, while absent in compact-dwarf plants. Linkage analysis showed that the markers OPF04700 and OPA091375 were located 8.4±0.03 cM and 9.6±0.032 cM, respectively away from the plant type gene locus. The markers were validated in 15 genotypes with open-tall plant type. The utilization of the marker and the dwarfing gene is discussed.

Keywords: Cajanus cajan, compact-dwarf, pigeonpea, plant type, marker, RAPD

 

 

Indian Journal of Biotechnology

Vol 9, January 2010, pp 64-68

Investigation on relative genome sizes and ploidy levels of Darjeeling-Himalayan Rhododendron species using flow cytometer

 

Kalyan Kumar De, Aniruddha Saha1*, Ranju Tamang and Bilok Sharma

Post Graduate Department of Botany, Darjeeling Government College, Darjeeling 734 101, India

1Department of Botany, University of North Bengal, PO-NBU, Darjeeling 735 013, India

Received 28 November 2008; revised 15 April 2009; accepted 2 July 2009

The relative 2C genome size (in pg), ploidy level (X) and total number of base pair (in Mbp) of ten threatened, rare and endangered Indian Rhododendron species of Darjeeling Hills (eastern Himalaya) were determined by using flow cytometer. Out of the 10 species, 9 were diploid and their genome sizes (2C) ranged from 1.30 to 1.51 pg. But in one species, i.e., R. niveum, the value of genome size was significantly high, i.e., 4.27 pg, appeared to be a natural hexaploid (6X). The cytometric genomic data indicating the hexaploid nature of R. niveum can be correlated with some phenotypic characters, specifically the size of leaf, size of flower and number of flower in truss. The reported phenotypic character of diploid species of R. niveum reveals that leaf size varies from 9-12 cm in length and 3.5-5.5 cm in breadth, individual flower size varies from 2-2.5 cm in length and 2-2.5 cm in diameter and number of flower per truss is 18-23 in diploid species. However, the findings on the same phenotypic characters of hexaploid R. niveum in the present study showed some differences from the diploid one. The leaf size of hexaploid R. niveum varies from 15-20 cm in length and 5-7 cm in breadth, individual flower size varies from 3.5-4 cm in length and 3.5 cm in diameter and number of flower per truss is 25-30. Therefore, these differences in floral traits and leaf size may be direct result of polyploidization itself. Thus, the morphological traits and the genomic information of Rhododendron species may serve as a valuable database for Indian Rhododendron breeders.

Keywords: Flow cytometer, genome size, Rhododendron

 

 

Indian Journal of Biotechnology

Vol 9, January 2010, pp 69-73

 

Evaluation and optimization of DNA extraction method for Dalbergia sissoo leaf

H S Ginwal* and Shalini Singh Maurya

Division of Genetics and Tree Propagation, Forest Research Institute, Kaulagarh Road, Dehradun 248 195, India

Received 23 December 2008; revised 15 June 2009; accepted 28 August 2009

A modified protocol for Dalbergia sissoo genomic DNA isolation has been optimized based on a cetyl trimethyl ammonium bromide (CTAB) method, described for other forest species. Leaves obtained from macro-propagated clones and mature trees of D. sissoo were tested. The method involves mortar grinding of tissue, a modified CTAB extraction buffer incorporating high salt concentrations, polyvinyl pyrrolidone and successive isoamyl alcohol-chloroform extractions with modified temperature conditions. The modification involved the use of doubled concentration of polyvinyl pyrrolidone (4% instead of 2%), increased incubation time with extraction buffer (40 min instead of 30 min), use of freshly prepared CTAB buffer and increased timing of washing of DNA pellet with wash buffer (45 min instead of 30 min). The yield was approximately 100 to 400 µg DNA per 100 mg of leaf tissue. The genomic DNA obtained by this method was suitable to be used in RAPD and ISSR analysis. This extraction method would allow the molecular analysis of DNA from different clones of D. sissoo.

Keywords: CTAB, Dalbergia sissoo, DNA extraction, forest tree, RAPD, ISSR

 

 

Indian Journal of Biotechnology

Vol 9, January 2010, pp 74-79

 

Differentiation of petroleum hydrocarbon-degrading Pseudomonas spp.
based on lectin binding of cell extracts in an agglutination assay

Bhaben Tanti and Alak Kumar Buragohain*

Department of Molecular Biology and Biotechnology, Tezpur University, Tezpur 784 028, India

Received 18 August 2008 ; revised 23 April 2009; accepted 9 July 2009

Plant and animal lectins with various carbohydrate specificities were used to type 43 petroleum hydrocarbon-degrading Pseudomonas isolates and the type strain P. aeruginosa MTCC1034 in a microtiter plate assay. Initially, a panel of nine lectins with their known specificity was used. For optimal typing, pretreatment by washing bacteria with a low pH buffer to allow protein release, followed by proteolytic degradation to eliminate autoagglutination, was used. Lectin types of treated samples were stable and reproducible. No strain proved to be untypeable by this system. The species were clustered into 10 different lectin types on the basis of their lectin binding with specific bacterial cell surface carbohydrate moieties. The result of lectin typing reveals considerable intraspecific variations within the species.

Keywords: Agglutination assay, carbohydrate moieties, lectin typing, Pseudomonas spp.

 

 

Indian Journal of Biotechnology

Vol 9, January 2010, pp 80-86

 

Antibiotic resistance profile of halophilic microorganisms isolated from tannery effluent

Rakesh Ghosh*, Pijush Kanti Chattopadhyay, Budhhadeb Chattopadhyay and Debasish Pal

Government College of Engineering and Leather Technology, Salt Lake, Kolkata 700 098, India

Received 17 December 2007; revised 1 April 2009; accepted 15 June 2009

Halophiles are defined as organisms showing considerable growth at salt concentrations higher than 100 g L-1. Based on the halophilicity, halophiles can be broadly classified as slightly, moderately or extremely halophilic depending on their requirement for NaCl.  Halophilic microorganisms, metabolically diversified, comprising Archaea, Bacteria, and Eucarya, are found distributed all over the world in hypersaline environments including drained soak liquor and brine cured hides.  Plasmids mediating resistance to antimicrobial agents have been found in many halophilic bacteria examined so far. For the purpose of protection of salt cured hides, adequate knowledge and exposure related to characteristics of halophilic bacteria is very important as halophilic microorganisms secrete bacterial collagenases responsible for collagen damage in the form of ‘Red heat’. All the halophilic bacteria isolated from the drained soak liquor used in these experiments were proved to be motile, aerobic and extremely pleomorphic Gram negative organisms. The growth curve of the halophilic bacteria showed slower growth profile at 37°C compared to E. coli. Effective plasmid isolation further strengthened the antibiotic resistance of the halophiles. Analysis of drained soak liquor was followed to examine the related important features of the halophilic species. Optimum salinity of media and pleomorphic Gram-negative nature of halophiles were found as causative factors of insensitivity to antimicrobial agents (AMA). It was found that darkness and low temperature would resist ‘Red heat’ on hides.

Keywords: Antibiotic resistance, curing, Gram negative, halophiles, plasmid, red heat, tannery effluent

 

 

Indian Journal of Biotechnology

Vol 9, January 2010, pp 87-95

 

Production of fibrolytic enzymes in repeat-batch culture using immobilized zoospores of anaerobic rumen fungi

Manpal Sridhar* and Deepak Kumar

National Institute of Animal Nutrition and Physiology, Adugodi, Bangalore 560 030, India

Received 16 September 2008; revised 20 March 2009; accepted 2 June 2009

The zoospores of two isolates of polycentric rumen fungi, Orpinomyces NIANP 58 (isolated from the faeces of a buffalo) and Anaeromyces NIANP 115 (isolated from the rumen liquor of a cannalulated cow), were immobilized in sodium alginate solution for the production of three fibrolytic enzymes, viz. CMcase, xylanase and b-glucosidase in repeat-batch culture.  Enzyme activity was recorded initially (0 h) and after every 24 h up to 72 h of immobilization. CMCellulase activity of 8.5±1.205 units at 0 h in Orpinomyces showed a very steep increase in the activity (598.00±26.87), almost70-fold increase at 48 h of immobilization. This increase was reflected in the specific activity as well, while in case of Anaeromyces, an activity of 6.0±0.851 units at 0 h also showed a very steep increase in activity (322.46±1.054 units, recording a 50-fold increase at 48 h of immobilization. At 0 h, 28.22± 4.556 units of xylanase activity corresponding to a specific activity of 31.36±6.225 were obtained in the case of Orpinomyces, which increased steeply to 955.11±54.93 units at 48 h of immobilization. Thereafter, the activity declined to more or less that obtained at 24 h of immobilization though a high specific activity of 791.20±10.66 was recorded. High xylanase activities were obtained in case of Anaeromyces upon immobilization with activity increasing from 56.88 units at 0 h to as high as 1921 units at 72 h. Anaeromyces isolate yielded 3.85, 4.246, 8.03 and 23.19 units of b-glucosidase, whereas Orpinomyces isolate yielded 6.09, 14.12, 16.63 and 20.89 units of b-glucosidase at 0,24,48 and 72 h, respectively. The results clearly elucidate the feasibility of using the zoospores of anaerobic rumen fungi for large-scale production of these three enzymes, which have great potential in ruminant nutrition in the breakdown of fibrous feeds and also various industrial applications.

Keywords: Anaerobic fungi, fibrolytic enzymes, batch culture, immobilization

 

 

Indian Journal of Biotechnology

Vol 9, January 2010, pp 96-100

 

Homology modeling and docking studies between HIV-1 protease and
carbamic acid

M Balakrishnan*, R C Srivastava and Mayank Pokhriyal

Bioinformatics Centre, Central Agricultural Research Institute, Port Blair 744 101, India

Received 16 April 2008; revised 11 December 2008 accepted 10 June 2009

HIV-I protease (HIV-I PR) is aspartic protease enzyme which is essential for the life-cycle of HIV retrovirus. Homology structural model and function relation of HIV-I PR have solved the structure of HIV-I proteases. We created a homology model of HIV-I PR and the 3-D structure as template using with ICMPro software. The ICMPro homology modeling algorithm has demonstrated excellent accuracy in blind predictions. Moreover, recent results show that ICMPro models built with as little as 35% identity can be accurate enough to be successfully used in receptor based rational drug design. The closest homologue with the highest sequence identity of 38.395% was selected as representative model using YASARA tools. The model was validated using protein structure checking tools such as PROCHEK for reliability. A total of two pockets were predicted by the software. Once the pockets were predicted, the ligand was subjected to docking reaction using the docking module of ICMPro software. Based on the RMSD and energy values, the best docking orientation was selected. The better RMSD value of docking is 0.0066288. This study will be used in broad screening of inhibitors of the protein and can be further implemented in future drug designing.

Keywords: Docking, energy minimization, HIV, homology modeling, protease, RMSD

 

 

Indian Journal of Biotechnology

Vol 9, January 2010, pp 101-105

 

Regeneration of multiple shoots from the callus cultures of
Macrotyloma uniflorum (Lam.) Verdc.

D H Tejavathi1*, V R Devaraj2, Savitha M Murthy1, P Anitha3 and R Nijagunaiah1

1Department of Botany, Jnanabharathi, Bangalore University, Bangalore 560 056, India
2Department of Biochemistry, Central College, Bangalore University, Bangalore 560 001, India
3Department of Botany, BMS College for Women, Basavanagudi, Bangalore 560 004, India

Received 13 March; revised 8 June 2009; accepted 21 August 2009

An effective protocol for the regeneration of multiple shoots from the callus, derived from shoot tip and cotyledonary node, was developed for the first time for Macrotyloma uniflorum (Lam.) Verdc. Shoot apices and cotyledonary nodes from 9-d-old aseptically grown seedlings were inoculated onto MS, L2 and MMS media supplemented with various growth regulators. Proliferation of about 17 shoots was obtained on AS supplemented MS medium after 16 d of culture, while about 14 and 20 shoots regenerated from the callus on MMS1+NAA (1.86 μM)+BAP (0.45 μM) and L2+IBA (0.45 μM)+BAP (0.11 μM), respectively after 30 d of culture. Regenerated shoots were rooted on MS+IAA (0.11 µM)+Kn (0.21 µM) with upto 6 roots per shoot. Rooted shoots were sequentially acclimatized by transferring them to sterile distilled water and tap water before planting in various pot mixtures. A maximum of 60% survival rate was noticed on a mixture of soil:sand:manure (1:1:1).

Keywords: Callus culture, histology, horse gram, Macrotyloma uniflorum, organogenesis