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Indian Journal of Biotechnology

 

 

ISSN:0972-5849

 

CODEN:IJBNAR 4(1) 1-164 (2005)

VOLUME 4

NUMBER 1

JANUARY 2005

 

 

 

CONTENTS

 

Reviews

 

Indian biotechnology: Challenges and opportunities - A clinician’s perspective

9

R D Lele

 

 

 

UlTRaSCAN-An algorithm for prediction of pattern(s) in untranslated regions of eukaryotic mRNAs

21

Rajeshwari Marikkannu & P Palanivelu

 

 

 

Post-harvest biotechnology of fruits with special reference to banana—Perspective and scope

39

K K Surendranathan

 

 

 

Cryopreservation of somatic embryos—An overview

47

Sonali Dixit Sharma

 

 

 

Papers

 

Amplified fragment length polymorphism (AFLP) analysis of genetic diversity in Indian mungbean [Vigna radiata (L.) Wilczek] cultivars

56

K V Bhat, S Lakhanpaul & S Chadha

 

 

 

Increased Fe-toxicity tolerance in rice calli and modulation in isozyme profiles

65

Bidhan Roy & Asit B Mandal

 

 

 

Transgenic cabbage (Brassica oleracea var. capitata) resistant to Diamondback moth (Plutella xylostella)

72

Anderson Paul, S R Sharma, T V S Sresty, Shantibala Devi, Suman Bala, P S Kumar, P Pardha Saradhi, Roger Frutos, I Altosaar & P Ananda Kumar

 

 

 

Plant regeneration from semi-mature zygotic embryos of Dalbergia sissoo Roxb.

78

Suresh Chand & Ajay Kumar Singh

 

 

Preclinical studies for gene therapy of head and neck cancers using the HSV-tk/GCV strategy

82

Archana S Wagle, Ganesh V Joshi, Kikkeri N Naresh & Rita Mulherkar

 

 

 

Cloning and expression of bovine (Bos indicus) interleukin-2 in Escherichia coli

88

Amar A Patil, Mohini Saini, Sameer Sharma, R B Bind & Praveen K Gupta

 

 

 

G and P genotyping of bovine group A rotaviruses in faecal samples of diarrhoeic calves by DIG-labelled probes

93

Minakshi, G Prasad, Y Malik & R Pandey

 

 

 

Seroepidemiological study of Plasmodium falciparum antigens for detection of malaria

100

M M Mya, R K Saxena & A Roy

 

 

 

Effect of excipients on product characteristics and structure of lyophilized lasota vaccine

106

S S Kadam, S S Pisal, J J Mane & M H Shah

 

 

 

Moisturizing efficiency of silk protein hydrolysate: Silk fibroin

115

A V Daithankar, M N Padamwar, S S Pisal, A R Paradkar & K R Mahadik

 

 

 

Quantitative structure activity relationship study of pyrazole ligands binding to estrogen receptor-α-selective agonists

122

Soumya Srivastava & S K Srivastava

 

 

 

Kinetic study of a low molecular weight protease from newly isolated Pseudomonas sp. using artificial neural network

127

Jayati Ray Dutta, Pranab Kumar Dutta & Rintu Banerjee

 

 

 

Purification and characterization of catechol 1, 2-dioxygenase of Pseudomonas fluorescens  for degradation of 4-chlorobenzoic acid

134

Preti Saxena & Indu Shekhar Thakur

 

 

 

Bioremediation of toxic metal ions using biomass of Aspergillus fumigatus from fermentative waste

139

K Rama Rao, K Rashmi, J Naveena Lavanya Latha & P Maruthi Mohan

 

 

 

Remazol brilliant blue reactive dye decolouration and mustard straw utilization by white rot fungi

144

S Vaithiyanathan, A S Mishra, R Prasad, M K Tripathi, A K Misra, O H Chaturvedi & R C Jakhmola

 

       

 

Short Communications

 

Potential industrial applications of yeast capable of fermenting high gravity cane molasses despite physiological stress

149

B K Bajaj, Vikas Taank & R L Thakur

 

 

 

Degradation of phorate by Azotobacter isolates

153

T A Kadam & L V Gangawane

 

 

 

Effect of bioinoculants on biomass productivity under agroforestry systems

156

Satyawati Sharma, Suman Kashyap & Padma Vasudevan

 

 

 

Author Index

 

 

 

Indian Journal of Biotechnology

Vol 4, January 2005, pp. 9-20

 

Indian biotechnology: Challenges and opportunities - A clinician’s perspective

R D Lele*

 

Biotechnology  has  provided  many  investigational  tools ¾ autoradiography  and  tracer  techniques,  X-ray crystallography, HPCL GC-MS and NMR spectroscopy, monoclonal antibodies, peptides, anti-sense oligonucleotides  and aptamers, to probe molecular events in the living human body including the brain. Knock-out and transgenic mouse models have revealed molecular mechanisms of many important human diseases such as hypertension, diabetes, atherosclerosis, neuro-degenerative disorders and cancer. New understanding of molecular mechanisms of disease leads to search for prevention as well as new therapies. The Human Genome Project has raised public hopes that doctors will detect and treat diseases like diabetes and cancer even before the symptoms begin, using medications that boost or counteract the effects of individual proteins, and they will know right from the start how to select the best medicine to suit each patient. Stem cell research has opened up the possibility of replacing or regenerating failing body parts with new tissues. Biologists and engineers working together in the fledgling field of tissue engineering are within the reach of constructing a living human heart patch. There is an urgent need for close and continuous interaction between the medical and biotechnology communities in India, to bring the full benefit of biotechnology for health care. A particular challenge and opportunity for Indian biotechnology is to build a golden triangle between ancient, experiential Indian medical wisdom, modern medicine and modern science, and validate the effectiveness of Ayurvedic drugs and practices in terms of current understanding of molecular biology and molecular pharmacology. For this we need imaginative thinking coupled with cutting-edge technology using the 100 available receptors, ion channels, transporters with their signaling molecules and enzymes along with their appropriate radiolabeled ligands. India with its vast population and great ethnic diversity is a gold mine for SNP studies, which have great implications for diagnosis, therapy as well as prevention.

 

Keywords: mAbs, antisens oligonucleotides, aptameters, cyclochrome P450, diabetes, human genome, peptides, stem cell

IPC Code: Int. Cl.7 A 61 K 35/74, 35/76, 38/00, 39/00; C 07 H; H 01 J

 

 

 

Indian Journal of Biotechnology

Vol 4, January 2005, pp 21-38

 

UlTRaSCAN - An algorithm for prediction of pattern(s) in untranslated regions of eukaryotic mRNAs

Rajeshwari Marikkannu1 and P Palanivelu2*

 

Many cis acting elements have been identified in the 3′ and 5′ untranslated regions (UTRs) of eukaryotic mRNAs. These cis acting elements are found to play vital roles in pre-mRNA processing, nucleo-cytoplasmic transport of processed mRNAs, determining the efficiency of translation of mRNAs and their stability and degradation in the cytoplasm. UTRScan utility has been used to identify these patterns in mRNAs. However, the UTRScan is not very sensitive and also not highly specific. It often generates many false positives as it scans the whole mRNA including the coding sequence. The authors have developed a new algorithm and an Internet based web application tool, named UlTRaSCAN, which overcomes these limitations and proved to be highly specific and sensitive in detecting patterns in UTRs. The new algorithm identifies these cis acting elements only in the 3′ and 5′ UTRs of eukaryotic mRNA/DNA sequences. The sensitivity is more than doubled and the specificity is increased, close to 100%. The UlTRaSCAN also minimized the false positives to almost 0% and the false negatives to large extent.

 

Keywords: eukaryotic mRNAs, 3′ untranslated region, 5′ untranslated region, UTRScan, UlTRaSCAN, UTR patterns

IPC Code: Int. Cl.7 C 12 N 15/10; G 06 F 15/00

 

 

 

 

Indian Journal of Biotechnology

Vol 4, January 2005, pp. 39-46

 

Post-harvest biotechnology of fruits with special reference to banana - Perspective and scope

K K Surendranathan*

 

Fruits, being highly nutritive, are important component of human diet but they possess very short post-harvest shelf life. As ripen, they become very soft and more prone to injuries, which makes them highly perishable. In India, over 30% of the annual produce is wasted due to spoilage. Hence, there is an urgent need to develop technologies to overcome post-harvest losses of fruits. Physiologists and biochemists attempted to extend the shelf life of fruits by different means though the results were not very satisfying. It was demonstrated recently that a judicious dose of g-irradiation (0.1-0.5 kGy) could enhance the shelf life to fruits by about a week to a fortnight, which could help in minimising the spoilage during storage and transportation. However, stringent quality controls have to be strictly followed to get the best results. Studies revealed that g-irradiation brings alterations/changes in metabolic pathways, which delay the production of essential precursors and energy required for ripening of fruits. Another strategy, to enhance the shelf life of fruits, could be adopted through regulation of endogenous ethylene production. Most recent studies have shown that it could be achieved by such genetically modified (GM) crops where gene expression of key enzymes responsible for ripening, like PG-ase, EFE and ACC-synthase, by means of antisense RNAs. However, adoption of this technology has so far been deterred due to apprehensions of safety issues associated with GM crops. An alternate method for prevention of spoilage of fruits as well as sustaining the interests in farmers could be the value addition of fruit commodities. This could be achieved by improving the conventional methods as well as development of non-conventional products of commercial interest. Nuclear Agriculture and Biotechnology Division, BARC has developed processes for the production of juice and powder from ripe banana, the largest produced and maximum wasted fruit, by creating an in-built mechanism to inactivate the pectin forming enzymes. With this process, over 60-80 % of the total moisture of the fruit is extracted out as juice. The commercially available variety, ‘Harichal’ (Mumbai kela) could give juice 550-640 ml /kg pulp. It is also demonstrated that a number of products of commercial significance, like banana nectar, carbonated juice and wine, from banana juice and biscuits, cakes, milkshakes, etc. from banana powder could be developed.

 

Keywords: banana juice, banana products, fruit ripening, mechanism of delay in ripening, post-harvest biotechnology

IPC Code: Int. Cl.7 A 23 L; C 12 N 15/01

 

 

 

Indian Journal of Biotechnology

Vol 4, January 2005, pp 47-55

 

Cryopreservation of somatic embryos - An overview

Sonali Dixit Sharma

 

Development of methods for long-term conservation of somatic embryos is important because they are potential explants for manipulations and conservation of in vitro maintained germplasm. This review discusses about the cryopreservation of somatic embryos with special reference to techniques involved and genetic stability of cryopreserved germplasm.

 

Keywords: cryopreservation, genetic stability, somatic embryos

IPC Code: Int. Cl. 7 A 23 B 7/024, 7/055, 9/10

 

 

 

Indian Journal of Biotechnology

Vol 4, January 2005, pp. 56-64

 

Amplified fragment length polymorphism (AFLP) analysis of genetic diversity in Indian mungbean [Vigna radiata (L.) Wilczek] cultivars

K V Bhat*, S Lakhanpaul and S Chadha

 

Released cultivars and improved lines of mungbean [Vigna radiata (L.) Wilczek] were subjected to AFLP (amplified fragment length polymorphism) analysis to test its usefulness and also to have an assessment of the genetic diversity and relationships among the cultivars. Relative efficiency of the primers, having three (+3) vs. two (+2) selective nucleotides, was also tested for detecting polymorphism. A total of 731 amplification products were obtained in the 27 cultivars with twelve primer pairs. Higher percent polymorphism was obtained with +3 than with +2 primers, though the number of amplification products was much higher with +2 primers. Consequently, higher average similarity coefficient (0.849) was obtained with +2 primers in comparison to +3 primers (0.751). Overall, a narrow genetic diversity (0.681-0.925) was recorded among the cultivars analysed. Distinct clusters were formed in the dendrogram with some variations in the constituents when data from +3 primers alone was compared with that from +2 primers. Principle coordinates analysis supports the results of UPGMA, as there was general agreement between the clustering patterns in both the analyses. The ‘Eigen’ vectors analysis indicated that the contributions of the first three factors were 12.49, 9.44 and 6.71, respectively. Twenty-seven factors were required to explain the total variation observed. Narrow genetic base observed is likely to be due to the use of limited material in the development of the cultivars analysed.

Keywords: AFLP, diversity analysis, green gram, mungbean, Vigna radiata

IPC Code: Int. Cl.7 C 12 N 15/10

 

 

 

Indian Journal of Biotechnology

Vol 4, January 2005, pp. 65-71

 

Increased Fe-toxicity tolerance in rice calli and modulation in isozyme profiles

Bidhan Roy* and Asit B Mandal

 

Seed derived calli of rice cultivars, ‘IR72’ (susceptible) and ‘C14-8’ (tolerant) were screened in vitro under increasing levels (50, 100, 200 or 400 ppm) of Fe-toxicity and profiled for isozymes, viz. esterase, peroxidase, malate dehydrogenase and lactate dehydrogenase, to assess their involvement in Fe-toxicity tolerance. In vitro screening showed the detrimental effect of hifher concentration on plantlet regeneration. However, a few calli survived on stressed medium and regenerated plantlets. Cultivar ‘C14-8’ showed higher degree of tolerance than ‘IR72’. Prominent differences including changes in band activity/intensity, mobility/shift and number of polymorphic loci with respect of Fe-toxicity were evident. The activity of esterase, malate dehydrogenase and lactate dehydrogenase decreased in Fe-stressed medium in ‘C14-8’. Decreased activity indicates gradual degradation of these enzymes or their structural modification under increased Fe-toxicity levels. On the other hand, the activity of these enzymes increased in the ‘IR72’ under stressed medium. However, the activity of peroxidase remained almost unaltered in ‘C14-8’ across the stress gradient. Few bands disappeared or newly appeared in the stressed medium as compared to control. This may be due to activation or inactivation of diverse domains in the genome and may be involved in governing Fe-toxicity tolerance. Whereas, a few bands remained unaltered across the stress gradient, and these may be used as biochemical marker for selection of tolerant plants against iron toxicity.

 

Keywords: Fe-toxicity tolerance, isozyme analysis, rice

IPC Code: Int. Cl.7 A 01 H 5/00; C 12 N 9/00

 

 

 

Indian Journal of Biotechnology

Vol 4, January 2005, pp. 72-77

 

Transgenic cabbage (Brassica oleracea var. capitata) resistant to Diamondback moth (Plutella xylostella)

Anderson Paul1, S R Sharma2, T V S Sresty3, Shantibala Devi 1, Suman Bala1, P S Kumar1, P Pardha Saradhi4, Roger Frutos5, I Altosaar6 and P Ananda Kumar

 

A synthetic fusion gene of Bacillus thuringiensis encoding a translational fusion product of Cry1B and Cry1Ab δ-endotoxins was transferred to a tropical cabbage breeding line by Agrobacterium-mediated transformation. Selection of transformants was carried out on media containing kanamycin. Polymerase chain reaction (PCR) analysis revealed that twelve of the putative transformants contained the transgene. Insect bioassays carried out with the leaves of PCR-positive plants and neonate larvae of Diamondback moth (DBM) showed that one of the transgenic plants was completely resistant to repeated infestation by the larvae. Southern hybridization confirmed gene integration in the DBM-resistant plant. Double-antibody sandwich Enzyme-Linked Immunosorbant Assay (ELISA) analysis revealed accumulation of fusion protein up to 0.16% of total soluble protein in the leaves of the transgenic plants. Progeny (T1 generation) of the selfed transgenic plants were analyzed for the transgene segregation and insect protection. These studies clearly demonstrated the efficacy of Cry1B-Cry1Ab fusion protein to confer protection to cabbage against DBM infestation. The transgenic cabbage plants will serve as a good system to study the role of gene pyramiding in resistance management strategies intended to prevent evolution of resistance in DBM.

 

Keywords: Bacillus thuringiensis, insecticidal protein genes, cry1Ab, cry1B, cabbage, Diamondback moth, transgenic plants

IPC Code: Int. Cl.7 C 12 N 15/00, 15/09, 15/11

 

 

 

Indian Journal of Biotechnology

Vol 4, January 2005, pp 78-81

 

Plant regeneration from semi-mature zygotic embryos of Dalbergia sissoo Roxb.

Suresh Chand* and Ajay Kumar Singh

 

Plant regeneration from callus cultures derived from semi-mature zygotic embryos of Dalbergia sissoo Roxb. was achieved. Callus formation occurred on Murashige and Skoog (1962) medium supplemented with 2.26-13.57 mM 2,4-D in combination with 0.46 and 1.16 mM Kn. Maximum response for callus formation was 78.3% on MS medium containing 9.04 mM 2,4-D and 1.16 mM Kn. Shoot regeneration occurred when calli clumps were transferred to MS medium enriched with 2.22-13.32 mM BAP and 1.34 mM NAA. Maximum response (45%) for shoot regeneration was achieved when calli clumps were transferred to MS medium supplemented with 8.88 mM BAP and 1.34 mM NAA. Average number of shoots per callus clump was 7.5 after 15 weeks of culture. In vitro regenerated shoots were rooted on ½ MS medium containing 1.23 mM IBA. Rooted plantlets transferred to pots containing autoclaved peat moss, soil, and compost mixture (1:1:1) were successfully established in pots.

 

Keywords: Dalbergia sissoo, plant regeneration, semi-mature zygotic embryos

IPC Code: Int. Cl.7 A 01 H 5/00, 5/10

 

 

 

Indian Journal of Biotechnology

Vol 4, January 2005, pp. 82-87

 

Preclinical studies for gene therapy of head and neck cancers using the HSV-tk/GCV strategy

Archana S Wagle1, Ganesh V Joshi1, Kikkeri N Naresh2 and  Rita Mulherkar1*

 

The present study has used the suicide gene Herpes Simplex Virus–thymidine kinase (HSV-tk) and ganciclovir (GCV) strategy. A 2 kb sequence of HSV-tk was subcloned into the retroviral vector resulting in a recombinant vector, LTKSN, which was transfected into a packaging cell-line, PA317 and selected on G418. The highest expressing clone, PTK-16 was used for in vitro experiments. Intra-tumoural injections of PTK-16 followed by GCV treatment, in a HNSCC xenograft model in nude mice, showed a significant reduction in the viable tumour volume (p=0.009). These results will form the basis for future clinical trials in HNSCC.

 

Keywords: gene therapy, HSV-tk, xenograft, nude mice, ganciclovir, head and neck cancer

IPC Code: Int. Cl.7 C 12 N 5/16, 15/12, 15/28

 

 

 

Indian Journal of Biotechnology

Vol 4, January 2005, pp. 88-92

 

Cloning and expression of bovine (Bos indicus) interleukin-2 in Escherichia coli

Amar A Patil1, Mohini Saini2, Sameer Sharma1, R B Bind1 and Praveen K Gupta1*

 

The gene for interleukin-2 (IL-2) was amplified from cDNA pool prepared from ConA-stimulated peripheral blood mononuclear cells (PBMCs) isolated from Indian cattle (Bos indicus). The amplified IL-2 gene was cloned and nucleotide sequences were determined. Homology comparison of nucleotide and predicted amino acid sequences revealed similarity of sequence and conservation of crucial amino acids with exotic cattle (B. taurus). The coding sequence of IL-2 (without its own signal sequence) was subsequently expressed as fusion protein with polyhistidine fusion tag in Escherichia coli, using prokaryotic expression vector. The expressed protein was present as insoluble inclusion bodies. The recombinant protein was solubilized with urea and purified using Ni-agarose affinity chromatography. The purified recombinant IL-2 was characterized in SDS-PAGE and in western blotting.

 

Keywords: Bos indicus, cDNA sequence, expression, interleukin-2, prokaryotic, recombinant

IPC Code: Int. Cl.7 C 12 N 15/26

 

 

Indian Journal of Biotechnology

Vol 4, January 2005, pp 93-99

 

G and P genotyping of bovine group A rotaviruses in faecal samples of diarrhoeic calves by DIG-labelled probes

Minakshi1, G Prasad1*, Y Malik3 and R Pandey2

 

The nucleic acid probes specific to genome segments 9 and 4 of bovine group A rotaviruses (BRVs) were developed for detection and determination of the G (G6 and G10) and P (P1, P5 and P11) types in field samples of diarrhoeic bovine calves. The type specific DNA probes were prepared by polymerase chain reaction amplification of hyper divergent regions of VP7 and VP4 genes from three genotypically distinct (G or P types) reference strains of BRV, viz. B223 (G10P11), NCDV (G6P1) and UK (G6P5), labelled with Digoxigenin (DIG). These G and P genotype specific DIG-labelled probes were evaluated for genotyping of rotaviruses of bovine origin directly in faecal samples. Of 308 calf diarrhoeic samples, 138 (44.81 %) were positive for BRV infection in nucleic acid hybridization assay, using VP7 and VP4 gene specific probes independently. Of probe positive samples (138), 112 were typeable by both G and P typing nested probes; 109 samples were typed as P11 (97.32%) and 3 as P1 (2.68%), and 97 as G10 (86.61%) and 15 as G6 (13.39 %). Surprisingly, none of the samples was typed as P5, indicating non-occurrence of P5 genotype among bovine rotaviruses in Haryana and adjoining areas of India. Furthermore, G10P11 was found to be the most prevalent combination among field rotavirus strains examined; whereas, G10P5 and G10P1 combinations were not reported in any of the samples.

 

Key words: BRV, DIG probes, dot-blot hybridisation, RT-PCR, G and P genotyping

IPC Code: Int. Cl.7 C 12 N 15/46

 

 

 

Indian Journal of Biotechnology

Vol 4, January 2005, pp. 100-105

 

Seroepidemiological study of Plasmodium falciparum antigens for detection of malaria

 M M Mya1, R K Saxena1* and A Roy2

 

Attempt has been made to develop a simple and cost-effective diagnostic method for the detection of malaria in field conditions. A new type of Plasmodium falciparum antigen (PSJ-M strain) from in vitro culture supernatant has been isolated. Purification and chemical analysis of the antigen showed that it is a glycophospholipid (GPL1), which contains mannose, xylose, glucose and galactose (3:3:1.5:1.5) in sugar moiety but its structure is devoid of inositol sugar and amino acids. Sensitivity and specificity properties of GPL1, when compared with existing P. falciparum antigens by Laser light scattering immunoassay (LIA) and Enzyme linked immunosorbent assay (ELISA), showed that the antigen has a very high sensitivity for detection of P. falciparum malaria antibodies; also specificity percentage was found to be 98%. The serological properties of GPL1 antigen have also been evaluated in endemic and non-endemic areas of different regions of the country. A simple, economical and handy malaria detection immunosensor (MDI) has been designed and developed for field areas. Laboratory and field trials for detection of P. falciparum malaria by MDI showed highly encouraging and good results. Data obtained by MDI from field areas, when compared in laboratory by LIA and ELISA techniques, showed that MDI method had better diagnostic ability and efficacy for detection of P. falciparum malaria (94-98%). Results of the study also suggest that GPL1 antigen and MDI can be used for detection and prevention of P. falciparum malaria.

 

Keywords: antigen, ELISA, field study, glycophospholipid, LIA, MDI, sensitivity, specificity

IPC Code: Int.Cl.7 A 61 K 39/002; G 01 N 33/53

 

 

 

Indian Journal of Biotechnology

Vol 4, January 2005, pp. 106-114

 

Effect of excipients on product characteristics and structure of lyophilized lasota vaccine

S S Kadam, S S Pisal*, J J Mane and M H Shah

 

The research was aimed to minimize product defects and lyophilization-induced denaturation of lasota vaccine. Sugars alone produced coherent cake but failed to protect virus during lyophilization. Maillard cake browning due to denaturation was evident in N,Z amine products. Polymers, PF-127 and PVP K-90 were able to produce porous cake structure and thus showed efficient water removal. DSC curves reveal endothermic melting corresponding to partial crystallization of PEG-6000 and PF-127, while remaining plugs were amorphous. The X-ray diffraction confirms the DSC findings. Aqueous IR of harvest revealed symmetrical α-helix of virus. Loss of alpha helix in lasota products is indicated by the decreased absorbance of 1654 cm-1 band. The qualitative comparisons of α-helix region in aqueous IR spectra are in correlation with antibody titer. Trehalose with most excipients gave better titer than sucrose. The stable glass matrix with arrested molecular mobility prevented unfolding of lasota. PF-127 was better stabilizer than PEG-6000 but both crystallized during freezing. Trehalse-PVP K-90 showed optimum product characteristics and maximum protection to antigen.

 

Keywords: additives, antibody titer, DSC, FTIR, lasota, lyophilization, XRPD

IPC Code: Int. Cl.7 A 23 B 7/024; A 61 K 39/155

 

 

 

Indian Journal of Biotechnology

Vol 4, January 2005, pp 115-121

 

Moisturizing efficiency of silk protein hydrolysate: Silk fibroin

A V Daithankar, M N Padamwar, S S Pisal*, A R Paradkar and K R Mahadik

 

A biomimetic approach of composition and natural function of natural moisturizing factor (NMF) with the amino acid content of silk fibroin was advantageously used to reconstruct the skin moisturizing system. The isolation of silk hydrolysate with water and sodium chloride treatment was complete in one hour. Lithium ion from LiBr effectively penetrated crystal domains of fibroin and gave desired solubility. Silk fibroin from Bombyx mori cocoons was non-allergic and biocompatible in skin and rabbit eye tests. The concentration dependent moisturizing efficacy of fibroin (1-5% w/v) in solution and cream form has been demonstrated by TEWL in vitro and in volunteers. As compared to dry and normal skin the fibroin containing cream revealed increased substantivity. The increased hydroxproline content was responsible for retaining higher moisture in the skin. This in turn maintained the skin in soft and supple state. The significant drop in impedance was observed within 1 hr of the application of fibroin and the effect was sustained for more than 6 hrs. Thus, increased hydration level in stratum corneum was achieved by fibroin treatment. The SEM of fibroin treated skin replicas showed a desired attribute of soft, smooth skin texture and improved flexibility. The increased state of hydration caused interdigitating of cell edges as evident in microphotographs. The rapid and sustained moisturizing efficiency observed with silk fibroin was well substantiated by the results of skin substantivity and impedance tests.

 

Keywords: silk fibroin, natural moisturizer, in vivo, TEWL, SEM, impedance

IPC Code: Int. Cl.7 A 61 K 7/40, 7/48

 

 

 

Indian Journal of Biotechnology

Vol 4, January 2005, pp 122-126

 

Quantitative structure activity relationship study of pyrazole ligands binding to estrogen receptor-α selective agonists

Soumya Srivastava1 and S K Srivastava2*

 

A quantitative structure activity relationship (QSAR) study on tetrasubstituted pyrazoles as high affinity ligands for the estrogen receptor (both ERα and ERβ subtypes) have been performed using various combinations of hydrophobic (MlogP), steric (MR) and electronic (Xeq) descriptors. The regression analysis of the data has shown better results in multiparametric regressions upon introduction of dummy parameters (indicator variables). The results suggest that the binding affinity for ERα and ERβ subtypes in tetrasubstituted pyrazoles is largely enhanced by the negative coefficient of MlogP and positive coefficient of MR descriptors.

Keywords: quantitative structure - activity relationship, estrogen receptors, tetrasubstituted pyrazoles

IPC code: Int. Cl.7 A 61 K 38/00; C 07 D 231/00

 

 

 

Indian Journal of Biotechnology

Vol 4, January 2005, pp 127-133

 

Kinetic study of a low molecular weight protease from newly isolated Pseudomonas sp. using artificial neural network

Jayati Ray Dutta1, Pranab Kumar Dutta2 and Rintu Banerjee1*

 

A soil isolate, identified as Pseudomonas sp. produced an extra-cellular protease enzyme of 14.4 kDa molecular weight. The kinetic properties of the purified fraction of the bacterial protease were studied experimentally and the rate of casein hydrolysis was predicted by a model based on artificial neural network. The various kinetic factors studied were incubation time, initial enzyme concentration, initial substrate concentration, pH and temperature. The prediction error in simulating casein hydrolysis was less than 1%.

 

Keywords: Pseudomonas sp., protease, kinetics, simulation, artificial neural network

IPC Code: Int. Cl.7 A 10 N 63/02; C 12 N 9/52

 

 

 

Indian Journal of Biotechnology

Vol 4, January 2005, pp. 134-138

 

Purification and characterization of catechol 1,2-dioxygenase of Pseudomonas fluorescens for degradation of 4-chlorobenzoic acid

Preti Saxena1and Indu Shekhar Thakur2*

The degradation of 4-chlorobenzoic acid (4-CBA) by Pseudomonas fluorescens IST8 was determined. The degradation of 4-CBA proceeded through an oxidative route to yield ortho ring cleavage enzyme, catechol 1, 2-dioxygenase. The cell free extract fractionated by DEAE-cellulose ion-exchange and gel filtration chromatography showed two different fractions of catechol 1,2-dioxygenase with an expected molecular weight of 62 and 48 kDa, respectively. The catechol 1, 2-dioxygenase in the fractions I and II was purified to about 22.3 and 36.5 fold by using purification steps. The pH and temperature optima for enzyme activity were 6.5 and 25°C, respectively. The purified protein on SDS-polyacrylamide gel electrophoresis showed molecular weights of 28 and 24 kDa, indicating dimeric nature of the enzyme.

Keywords:   catechol-1,2-dioxygenase, 4-chlorobenzoic acid, chlorocatechol, DEAE-cellulose, gel filtration, SDS-polyacrylamide gel electrophoresis

IPC Code: Int. Cl.7 A 01 N 29/00, 63/00; C 07 C 63/10

 

 

 

Indian Journal of Biotechnology

Vol 4, January 2005, pp 139-143

 

Bioremediation of toxic metal ions using biomass of Aspergillus fumigatus from fermentative waste

K Rama Rao, K Rashmi, J Naveena Lavanya Latha and P Maruthi Mohan*

 

Dried, nonliving, granulated biomass of Aspergillus fumigatus from fermentation industry was used for the removal of Cd2+, Co2+, Cu2+ and Ni2+ from solutions. Sorption studies showed sequestration (70-90%) of Cd2+ from solutions (0.1-4 mM). However, with increase in concentration, Cd2+ sorption efficiency decreased due to saturation of the biosorbent. Cu2+ binds most efficiently (72%) to the biosorbent followed by Cd2+ (61%), Co2+ (49%) and Ni2+ (37%). Metal removal from solutions containing a mixture of metal ions (Cd2+, Cu2+, Co2+, and Ni2+), which reflects the features of the polluted wastewaters and industrial effluents, was also efficient (90%) at lower concentrations (0.1 mM each). At higher concentrations (5 mM to 25 mM), Cu2+ removal was predominant (>70%) over other ions. The biosorbent was reusuable up to 5 cycles with a 50% loss of initial Cd2+ binding capacity. However, a significant loss of Cd2+ binding capacity was observed when biosorbent was immobilized in polyvinyl foam. Infrared spectra of the biosorbent preparation showed the involvement of alcohol/amine (OH/NH2) and CH-OH functional groups in metal binding. The present studies suggest that fungal biomass, a waste from fermentative industry, has the potential for removal/recovery of toxic metal ions from aqueous solutions.

 

Keywords: Aspergillus fumigatus, binding capacity, bioremediation, biosorption, metals

IPC Code: Int. Cl.7 C 12 N 1/04, 1/24

 

 

 

Indian Journal of Biotechnology

Vol 4, January 2005, pp 144-148

 

Remazol brilliant blue reactive dye decolouration and mustard straw utilization by white rot fungi

S Vaithiyanathan*, A S Mishra, R Prasad, M K Tripathi, A K Misra, O H Chaturvedi and R C Jakhmola

 

The remazol brilliant blue reactive (RBBR) dye decolouration and mustard (Brassica campestris) straw (MuS) nutrient utilization by various white rot fungi, viz. Phanerochaete chrysosporium, Ganoderma applanatum, G. lucidum, Pleurotus ostreatus, P. sajorcaju, Polyporus arcularius, P. versicolor, P. adustus, P. sanguineus-970, P. sanguineus-154, Trametes hirsuta, Parva mentocela and Longyites strata, were studied. The visual detection of decolouration was found compleimentary to its quantitative estimation. P. sanguineus-970 showed the highest RBBR dye decolouration in agar as well as in broth medium (74%). The dry matter utilization of MuS was low (20-27%) by all the white rot fungi. However, they preferentially degraded lignin (18-42%) in comparison to cellulose (0-12%). The highest lignin utilization was shown by P. chrysosporium. Whereas, P. sanguineus-154 showed both higher RBBR dye decolouration (66.5%) and lignin degradation (33%) and could be used for modification of MuS as animal feed.

 

Keywords: decolouration, mustard straw, RBBR dye, white rot fungi

IPC Code: Int. Cl.7 C 12 N 1/14, 1/22

 

 

 

Indian Journal of Biotechnology

Vol 4, January 2005, pp. 149-152

 

Potential industrial applications of yeast capable of fermenting high gravity cane molasses despite physiological stress

B K Bajaj1*, Vikas Taank2 and R L Thakur2

 

Two Saccharomyces cerevisiae isolates were studied for their ability to ferment cane molasses of high gravity. Supplements like soybean meal (SM), groundnut meal (GM) or castor oil meal (CM) (@2.5%) were found to be quite effective in enabling the yeast to ferment molasses of high gravity (35-40° brix). Ethanol production efficiency was increased by 45-50% with supplementation of any of these additives to concentrated molasses. The viability of yeast also improved by 24-25% in high gravity molasses. In concentrated worts with no supplement, trehalose content of yeast was increased significantly but little increase was reported in medium supplemented with SM, GM or CM, indicating the stress relieving effect of these supplements. Glycerol content did not vary with increase in the concentration of sugars in the medium. There was also no effect on invertase activity of yeast while fermenting the concentrated molasses worts.

 

Keywords: ethanol production efficiency, fermentation, glycerol, high gravity molasses, osmotic stress, Saccharomyces cerevisiae, trehalose

IPC Code: Int. Cl.7 C 12 N 1/18, 1/22

 

 

 

Indian Journal of Biotechnology

Vol 4, January 2005, pp. 153-155

 

Degradation of phorate by Azotobacter isolates

T A Kadam and L V Gangawane*

 

Of 44 Azotobacter isolates from the soils of Maharashtra, 29 belonged to A. chroococcum and other were A. vinelandii. All A. chroococcum isolates were efficient in N fixation when tested by acetylene reduction test and also in cotton cv NHH-44. However, they showed variation to phorate tolerance. The maximum tolerance (150 mg/ml) was shown by ACI-17, ACI-18 and ACI-28 isolates. These isolates also showed maximum phorate degradation. TLC and HPLC studies indicated the presence of O,O-diethyl S-{(ethyl sulphonyl) methyl} phosphorothioate, O,O-diethyl S-{(ethyl sulphinyl) methyl} phosphorothioate and diethyl phosphoric acid as degradation products. Plasmid analysis indicated that the genes for phorate degradation might be located on the main chromosome and not on the plasmid.

 

Keywords: Azotobacter, N fixation, phorate, tolerance

IPC Code: Int. Cl.7 A 01 N 63/02

 

 

 

Indian Journal of Biotechnology

Vol 4, January 2005, pp. 156-160

 

Effect of bioinoculants on biomass productivity under agroforestry systems

Satyawati Sharma*, Suman Kashyap and Padma Vasudevan

 

Efficacy of native bioinoculants, viz. AM fungi and Azotobacter, separately as well as in combination was evaluated for enhancing biomass productivity of Morus alba, Populus deltoides, Psidium guajava and Leucaena leucocephala under different agroforestry models along with other plant species. The combination of all plant species tested was found to be favourable with respect to growth, yield and microbial population in soil. The combined inoculation of AM fungi and Azotobacter gave the best results. AM fungi were cultured on Zea mays, Trigonella foenum-graecum, Ricinus communis, Vigna radiata, Solanum melongena and Lycopersicon esculenum, wherein R. communis was found to be the best host plant.

 

Keywords: agroforestry, AM fungi, Azotobacter

IPC Code: Int. Cl.7 A 01 N 63/02, 63/04

 

 

AUTHOR INDEX

 

 

Altosaar I                           72

Ananda Kumar P                72

 

Bajaj B K                         149

Bala S                                72

Banerjee R                       127

Bhat K V                           56

Bind R B                            88

 

Chadha S                           56

Chand S                             78

Chaturvedi O H                144

 

Daithankar A V                115

Devi S                                72

Dutta J R                          127

Dutta P K                         127

 

Frutos R                             72

 

Gangawane L V               153

Gupta P K                          88

 

Jakhmola R C                   144

Joshi G V                           82

 

Kadam S S                       106

Kadam T A                      153

Kashyap S                        156

Kumar P S                         72

 

Lakhanpaul S                  56

Latha J N L                  139

Lele R D                          9

 

Mahadik K R                115

Malik Y                          93

Mandal A B                   65

Mane J J                      106

Marikkannu R                 21

Maruthi Mohan P          139

Minakshi                        93

Mishra A S                   144

Misra A K                    144

Mulherkar R                   82

Mya M M                     100

 

Naresh K N                   82

 

Padamwar M N            115

Palanivelu P                   21

Pandey R                       93

Paradkar A R               115

Pardha Saradhi P            72

Patil A A                        88

Paul A                           72

Pisal S S                106, 115

Prasad G                        93

Prasad R                      144

Rama Rao K           139

Rashmi K                139

Roy A                     100

Roy B                       65

 

Saini M                     88

Saxena P                 134

Saxena R K             100

Shah M H                106

Sharma Sameer         88

Sharma Satyawati    156

Sharma S D              47

Sharma S R               72

Singh A K                 78

Sresty T V S             72

Srivastava S             122

Srivastava S K         122

Surendranathan K K  39

 

Taank V                  149

Thakur I S               134

Thakur R L              149

Tripathi M K            144

Vaithiyanathan S      144

Vasudevan P           156

Wagle A S                82