@article { ISI:000231841800033, title = {Optimization and scale up of production of alkaline protease from Conidiobolus coronatus}, journal = {Process Biochemistry}, volume = {40}, number = {9}, year = {2005}, month = {SEP}, pages = {3152-3158}, publisher = {ELSEVIER SCI LTD}, type = {Article}, address = {THE BOULEVARD, LANGFORD LANE, KIDLINGTON, OXFORD OX5 1GB, OXON, ENGLAND}, abstract = {

Alkaline protease secreted by Conidiobolus coronatus has been evaluated extensively in tanneries and finds application in pre-tanning operations in leather manufacture. It is important to produce the enzyme in inexpensive and optimized media on large scale for the process to be commercially viable. The present paper describes optimization of fermentation conditions in shake flasks and scale up of production to 100 L in fermentors. The production is complete in 2-3 days comparable to bacterial fermentations. The organism utilized several carbon sources such as starch, sucrose, lactose, glucose and fructose for production. Soyabean meal at an optimum concentration of 2-3\% was found to be best inducer. Diammonium hydrogen phosphate, casamino acids and Hi-media peptone gave activities comparable to yeast extract. Preservation and stabilization studies showed that glycerol conferred considerable stabilization at room temperature while ammonium sulphate precipitated enzyme at 0.9 saturation was best with stability up to 2 years even at room temperature. (c) 2005 Elsevier Ltd. All rights reserved.

}, keywords = {Alkaline Protease, Conidiobolus, Fermentation, Optimization, scale up}, issn = {1359-5113}, doi = {10.1016/j.procbio.2005.04.005}, author = {Laxman, Ryali Seeta and Sonawane, A. P. and More, Shivaji V. and Rao, B. S. and Rele, M. V. and Jogdand, Vitthal V. and Deshpande, V. V. and Rao, M. B.} } @article { ISI:000298662900014, title = {Immobilization of conidiobolus coronatus alkaline protease on waste fungal biomass}, journal = {Environmental Engineering and Management Journal}, volume = {10}, number = {11}, year = {2011}, month = {NOV}, pages = {1727-1732}, publisher = {GH ASACHI TECHNICAL UNIV IASI}, address = {71 MANGERON BLVD, IASI, 700050, ROMANIA}, abstract = {

Alkaline protease from Conidiobolus coronatus which is optimally active at pH 10 and 40 degrees C finds application in leather and detergent industries as well as for recovery of silver from waste photographic films. The protease was immobilized in Ca-alginate, polyacrylamide gel and alkali treated waste fungal biomass (ATWFB). ATWFB was found to be most suitable among the matrices tested. Glutaraldehyde marginally increased the binding to ATWFB. Binding of protease to ATWFB seems to be through adsorption as confirmed by FTIR spectra. Though temperature optima of free and immobilized proteases were identical, optimum pH of the immobilized enzyme shifted to 11 from 10. Temperature stability of the protease increased after immobilization. Immobilized protease could be reused 3 times with marginal loss in activity.

}, keywords = {Alkaline Protease, Conidiobolus coronatus, fungal biomass, Immobilization}, issn = {1582-9596}, author = {Shankar, Shiv and Laxman, Ryali Seeta} } @article { ISI:000287274000022, title = {Purification and characterization of an alkaline protease by a new strain of Beauveria sp}, journal = {Process Biochemistry}, volume = {46}, number = {2}, year = {2011}, month = {FEB}, pages = {579-585}, publisher = {ELSEVIER SCI LTD}, address = {THE BOULEVARD, LANGFORD LANE, KIDLINGTON, OXFORD OX5 1GB, OXON, ENGLAND}, abstract = {

A fungal culture isolated from animal dung was identified as a new strain of Beauveria sp MTCC 5184 based on 18S rDNA and ITS nucleotide sequence homology. The fungal isolate secretes alkaline protease active at pH 9 and 50 degrees C. The alkaline protease from Beauveria sp (BAP) was purified to homogeneity with 10.2-folds increase in specific activity and 38.6\% recovery. The molecular mass and isoelectric point of the protease were found to be 29 kDa and 9.3, respectively. The N-terminal sequence of the BAP showed only partial homology with subtilisin like proteases from other fungi. The enzyme was stable up to 40 degrees C and pH 3-11. The protease was inhibited by Cd(2+), Hg(2+) and Mn(2+). The activity was totally lost in the presence of 1 mM PMSF suggesting it to be a serine protease. The protease showed maximum activity with casein followed by haemoglobin and BSA. The purified protease is able to separate the endothelial cells and can be used in animal cell culture. (C) 2010 Elsevier Ltd. All rights reserved.

}, keywords = {Alkaline Protease, Animal cell culture, Beauveria sp, Sequence homology}, issn = {1359-5113}, doi = {10.1016/j.procbio.2010.10.013}, author = {Shankar, Shiv and Rao, Mala and Laxman, Ryali Seeta} } @article { ISI:000320099400002, title = {Enzymatic degumming of silk with microbial proteases}, journal = {Journal of Natural Fibers}, volume = {10}, number = {2}, year = {2013}, month = {JAN}, pages = {98-111}, publisher = {TAYLOR \& FRANCIS INC}, address = {325 CHESTNUT ST, SUITE 800, PHILADELPHIA, PA 19106 USA}, abstract = {

Degumming of Chinese bivoltine silk with alkaline proteases from various microbial sources was investigated and compared with commercial enzymes. Among the proteases tested, two fungal and two actinomycete proteases were promising, which showed weight loss similar to conventional method (19.58\% to 21.78\%). Conidiobolus brefeldianus and BOA-2 proteases were best enzymes, which showed weight loss similar to conventional method with low enzyme concentrations and in shorter time. No significant differences were found in tensile strength or elongation at break by enzymatic degumming indicating that there was no damage to the fiber. Scanning electron micrographs showed the sericin deposits were removed and the fibers were separated.

}, keywords = {Alkaline Protease, bivoltine, degumming, microbial, Silk, weight loss}, issn = {1544-0478}, doi = {10.1080/15440478.2012.761114}, author = {More, Shivaji V. and Khandelwal, Harish B. and Joseph, M. A. and Laxman, Ryali Seeta} } @article { ISI:000348102900047, title = {Biophysicochemical characterization of an alkaline protease from beauveria sp. MTCC 5184 with multiple applications}, journal = {Applied Biochemistry and Biotechnology}, volume = {175}, number = {1}, year = {2015}, month = {JAN}, pages = {589-602}, publisher = {HUMANA PRESS INC}, address = {999 RIVERVIEW DRIVE SUITE 208, TOTOWA, NJ 07512 USA}, abstract = {

This study illustrates the biophysicochemical properties of an alkaline protease, BAP (Beauveria sp. alkaline protease) from Beauveria sp. MTCC 5184. This protease exhibited maximum activity at 50 degrees C, pH 9.0, and stability in a broad pH range, in the presence of organic solvents, denaturants, as well as detergents. Wash performance studies revealed that BAP was able to remove blood clots/stains from blood-soaked cloth. Peptide mass fingerprinting results demonstrated partial homology of BAP with subtilisin-like proteinase. BAP showed catalytic activity against natural as well as synthetic substrates. Active site characterization of BAP confirmed the involvement of serine, tryptophan, and aspartic acid in catalytic activity. Detailed kinetic and thermodynamic studies of BAP demonstrated that the activation energy (Ea) for casein hydrolysis was 82.55 kJ/M, the specificity constant (Kcat/K-m), and the values of Delta G (change in Gibbs free energy) decreased with increase in temperature, whereas Delta H (change in enthalapy) and Delta S (change in entropy) were constant. The results of the present study indicate that BAP has potential for applications as detergent additive, in peptide synthesis, and in basic research.

}, keywords = {Active site, Alkaline Protease, Beauveria sp, Organic solvent, Substrate kinetics}, issn = {0273-2289}, doi = {10.1007/s12010-014-1314-3}, author = {Shankar, Shiv and Laxman, Ryali Seeta} } @article { ISI:000349853500010, title = {Eco-friendly enzymatic dehairing of skins and hides by C-brefeldianus protease}, journal = {Clean Technologies and Environmental Policy}, volume = {17}, number = {2}, year = {2015}, month = {FEB}, pages = {393-405}, publisher = {SPRINGER}, address = {233 SPRING ST, NEW YORK, NY 10013 USA}, abstract = {

Alkaline protease from Conidiobolus brefeldianus was efficient in unhairing various types of skins and hides. The crude protease preparation was active toward keratin-azure, elastin-orcin, azocasein, and azocoll, but did not show true collagenase activity. In addition, the crude enzyme exhibited other enzyme activities such as chondroitinase, laminarase, and chitinase. Complete hair removal of skin/hide by the protease achieved in 16-18 h. The dehaired pelt showed smooth and white appearance due to hair removal along with epidermal layer. In addition, the grain was clean and without damage in enzymatically dehaired pelts. The microscopic observation of the cross-section of dehaired goat skin and cow hide showed absence of epidermis, hair shaft with empty follicles. Enzymatic dehairing resulted in complete and uniform fiber opening in the dermis and corium region. Physical properties viz. tensile strength, elongation, and tear strength of dyed crust of enzymatically and conventionally dehaired pelts were comparable. Results were also validated on large scale with goat skins and cow hides.

}, keywords = {Alkaline Protease, Conidiobolus brefeldianus, Cow hides, Dehaired pelts, Enzymatic dehairing, Sheep skins}, issn = {1618-954X}, doi = {10.1007/s10098-014-0791-y}, author = {Khandelwal, Harish B. and More, Snehal V. and Kalal, K. M. and Laxman, Ryali Seeta} } @article { ISI:000352343600011, title = {Green synthesis of silver nanoribbons from waste X-ray films using alkaline protease}, journal = {Materials Express}, volume = {5}, number = {2}, year = {2015}, month = {APR}, pages = {165-170}, publisher = {AMER SCIENTIFIC PUBLISHERS}, address = {26650 THE OLD RD, STE 208, VALENCIA, CA 91381-0751 USA}, abstract = {

This work illustrates a facile approach to synthesize silver nanoribbons from waste X-ray films using alkaline protease from Beauveria sp. MTCC 5184 (BAP). The green synthesized nanoribbons have been characterized by UV-Vis spectroscopy, TEM, and FTIR. The UV-visible spectra gave maximum absorbance at 400 nm. TEM micrograph showed well-dispersed silver nanoribbons with an average length of 200-400 nm. FTIR result demonstrated the respective bands of the silver nanoribbons and proteins. XRD data revealed crystalline nature of silver nanoribbons dominated with (111) facets. In vitro antibacterial activity in terms of minimum inhibitory concentrations (MIC) and minimum bactericidal concentration (MBC) showed 8/16 mu g/mL (MIC/MBC) against Staphylococcus aureus ATCC 29213 and 4/8 mu g/mL (MIC/MBC) against Escherichia coli ATCC 25922. The green synthesized silver nanoribbons showed excellent biocompatibility up to 40 mu g/mL concentrations on mouse fibroblast cell line (L929).

}, keywords = {Alkaline Protease, Characterization, Cytotoxicity, Silver Nanoribbons, Waste X-ray Film}, issn = {2158-5849}, doi = {10.1166/mex.2015.1221}, author = {Shankar, Shiv and Prasad, R. G. S. V. and Selvakannan, P. R. and Jaiswal, Lily and Laxman, Ryali Seeta} } @article {44035, title = {Silk degumming and utilization of silk sericin by hydrolysis using alkaline protease from beauveria Sp. (MTCC 5184): a green approach}, journal = {Journal of Natural Fibers}, volume = {15}, year = {2017}, month = {JUL}, pages = {1-11}, type = {Article}, abstract = {Conventionally, degumming is carried out at 90{\textdegree}C--110{\textdegree}C temperature by boiling the raw silk with Marseilles soap and sodium bicarbonate which eventually requires a lot of water and energy. In this study, degumming of Chinese bivoltine raw silk fibres with alkaline protease produced by Beauveria sp. (MTCC 5184) is studied. Complete degumming was obtained in 45 min with 75 units of enzyme per gram of silk. Degumming was found to be optimal at 50{\textdegree}C and pH 9.0. Scanning electron micrographs showed that the sericin deposits were removed and the obtained fibres were clean, separated, had smooth feel with shine as compared to untreated fibres. Sericin isolated from silk cocoon (by-product which goes waste) was hydrolyzed with the same alkaline protease obtained from Beauveria sp. to get small molecular weight peptides. These peptides can be utilized further for cosmetic, pharmaceutical, and various industrial applications.}, keywords = {Alkaline Protease, bivoltine, degumming, Silk, weight loss}, issn = {1544-0478}, doi = {10.1080/15440478.2017.1330718}, author = {More, Snehal V. and Chavan, Sakalya and Prabhune, Asmita Ashutosh} }