• Analysis of Amino Acids
    At Celignis Analytical

Content coming soon

Publications on Amino Acids By The Celignis Team

V. P. Zambare and S. S. Nilegaonkar (2016) Proteases in Leather Processing, Industrial Biotechnology, Sustainable Production and Bioresource Utilization, Apple Academic Press, CRC Press

Link

V. P. Zambare, S. S. Nilegaonkar and P. P. Kanekar (2014) Scale up production of protease using Pseudomonas aeruginosa MCM B-327 and its detergent compatibility, Journal of Biochemical Technology 5(2): 698-707

Link

The Maximum Protease Activity Was Obtained From P. Aeruginosa MCM B-327 With Soybean Meal 1%, Tryptone 1%, Initial Medium PH 7, Agitation Rate 250 Rpm, Aeration Rate 0.75 Vvm And Fermentation Temperature 30 C, Under Submerged Fermentation Conditions (SmF). The Protease Productivity At 10 And 120L Fermenters Was Found To Be 16,021 And 9,975 UL-1h-1 Respectively. Kinetics Of Cell Growth Revealed That Specific Cell Growth Rate Was 0.025 H-1. Protease Was Active And Stable At Different PH, Temperatures, In Anionic, Cationic And Non-Ionic Detergent Additives, As Well As In Commercial Detergents. The Protease Exhibited Blood Stains Removing Performance Indicating Its Potential In Detergent Industry. The Dried Ammonium Sulphate Precipitated Protease Was Stable At Room Temperature For A Period Of One Year. The Protease Has Shown Properties Suitable For Its Application In Detergents. The Results Contribute To Basic Knowledge And Application Of Protease From P.Aeruginosa To Detergent Industry. The Studies Will Help To Optimize The Production Of This Protease For Biotechnological Applications.

V. P. Zambare, S. S. Nilegaonkar, P. P. Kanekar (2011) Production optimization and purification of a novel extracellular protease from Pseudomonas aeruginosa MCM B-327, New Biotechnology 28(2): 173-181

Link

The focus of this study was on production, purification and characterization of dehairing protease from Pseudomonas aeruginosa MCM B-327, isolated from vermicompost pit soil. Optimum protease activity, 395 U mL?1, was observed in the medium containing soybean meal and tryptone, at pH 7 and 30C. The crude enzyme exhibited dehairing activity. As compared to chemical method, enzymatic method of dehairing showed reduction in COD, TDS and TSS by 34.28%, 37.32% and 51.58%, respectively. Zymogram of crude enzyme on native-PAGE presented two bands with protease activity of molecular weights of 56 and 67 kDa. Both proteases showed dehairing activity. Out of these, 56 kDa protease (PA02) was purified 3.05-folds with 2.71% recovery. The enzyme was active in pH range 79 and temperature 2050C with optimum pH of 8 and temperature 35C. Moreover, the enzyme activity of PA02 protease was not strongly inhibited by specific inhibitor showing the novel nature of enzyme compared to serine, cysteine, aspartyl and metalloproteases. Kinetic studies indicated that substrate specificity of PA02 protease was towards various natural and synthetic proteolytic substrates but inactive against collagen and keratin. These findings suggest protease secreted by P. aeruginosa MCM B-327 may have application in dehairing for environment-friendly leather processing.

V. P. Zambare, S. S. Nilegaonkar, P. P. Kanekar (2011) Use of agroresidues for protease production and application in degelatinazation, Research Journal of BioTechnology 6(2): 62-65
Vasudeo Zambare (2011) Optimization of amylase production from Bacillus sp. using statistics based experimental design, Emirates Journal of Food and Agriculture 23(1): 37-47

Production of amylase under submerged fermentation Bacillus sp. was investigated using wheat bran, soybean meal and CaCO3 (WSC) medium. Response surface methodology (RSM) was used to evaluate the effect of the main variables, i.e., pH (11.35), temperature (35.16C) and inoculum size (2.95%) on amylase production by applying a full factorial central composite design (CCD). The mutual interaction between these variables resulted into 4.64 fold increase in amylase activity as compared to the non-optimized environmental factors in the basal medium.

Vasudeo Zambare (2010) Strain improvement of alkaline protease from Trichoderma reesei MTCC-3929 by physical and chemical mutagen, The IIOAB Journal 1(1): 25-28

Link

The purpose of the present investigation is to enhance alkaline protease production by subjecting indigenous protease producing strain Trichoderma reesei MTCC-3929 to improvement by random mutagenesis by ultra-violet (UV) irradiation and N-Methyl-N-nitro-N-nitroso guanidine (NTG) treatment. Mutants were screened as protease producers on the basis of zone of clearance on skimmed milk agar plates. UV-8 mutant showed 9 mm clear zone diameter and activities of 199.6 and 552.6 U/ml for submerged fermentation (Smf) and solid state fermentation (SSF), respectively. UV-8 further mutated by NTG to produced NTG-17 mutant with zone of clearance 13mm diameter. Compared to wild strain, NTG-17 mutant was found to produce 2.6 and 2.2-fold more activities in SmF and SSF, respectively. Thus these findings have more impact on enzyme economy for biotechnological applications of microbial proteases.

V. P. Zambare (2010) Optimization of nutritional parameters for extracellular protease production from Bacillus sp. using response surface resistance, International Journal of BioEngineering and Technology 1(1): 43-47

Link

The optimization of nutritional parameters and concentrations for the protease production by Bacillus sp. in submerged fermentation was carried out using response surface methodology (RSM) based on the central composite design (CCD). The design contains a total of 20 experimental trials containing starch, soybean meal and CaCO3 as model factors. The mutual interaction between these variables resulted into 1.48 fold increase in protease activity as compared to the mean observed response at zero level of all variables.

Vasudeo Zambare, Smita Nilegaonkar, Pradnya Kanekar (2010) Application of protease from Bacillus cereus MCM B-326 as a bating agent in leather processing, The IIOAB Journal 1(3): 18-21

Link

Laboratory scale experiments were carried out to test the efficiency of the extracellular protease from Bacillus cereus MCM B-326; cattle dung and commercial bate powder (ComBate) as bating agents on delimed buffalo hide. Protease treated pelt was free from scud and pigments, clean and fine grain, white, smooth and silkier with loosen fat. Histological sections of bated pelts showed greater opening up of collagen fibers with Bacillus protease. The studies indicated potential importance of Bacillus protease as effective bating agent in leather processing.

Vasudeo Zambare (2010) Purification and characterization of neutral serine protease from Bacillus sp., Asiatic Journal of Biotechnology Resources 3: 183-192

Link

A neutral protease was purified from Bacillus sp. to electrophoretic homogeneity by using ammonium sulphate precipitation and 2-step-column chromatography. The purified protease expressed its maximum activity 40 o C and pH value of 7. It was stable up to 10-40 o C for 30 min of incubation and retained 80 and 65% of its activity at 50 and 60 o C, respectively. Ions of Ca and Na and showed a stimulatory effect and ions of K and Fe had no effect, ions of Mg, Cu, Zn and Mn showed an inhibitory effect. Moreover, ions of Hg showed strong inhibitory effect on the purified protease activity. Neutral protease was found to be a serine protease and confirmed by enzyme inhibition using phenylmethylsulfonylfluoride (PMSF). The enzyme has high affinity towards casein followed by Bovine serum albumin (BSA) and gelatin. Molecular weight of the purified NP was found to be 35kDa on SDS-PAGE.

S. S. Nilegaonkar, V. P. Zambare, P. P. Kanekar, P. K. Dhakephalkar, S. S. Sarnaik, N. K. Chandrababu, Rama Rajaram, B. Ramanaiah, T. Ramasami, Y. K. Saikumari and P. Balaram (2007) A novel protease for industrial application, German Patent Patent NO. 102007013950.2

The present invention relates to an extracellular enzyme protease obtained by growing the culture of Pseudomonas aeruginosa MCM B-327 isolated from vermiculture pit soil and deposited in MTCC, IMTECH, Chandigarh with designation MTCC 5270, in production medium of pH 7.0; containing soybean meal and tryptone as raw materials, at 30 C. for 72 h. The organism was also able to produce protease using different agricultural products/byproducts as protein sources. The partially purified non-collagenolytic, calcium independent protease with molecular weight 60 kDa has activity in pH range of 6.0-11.0 and temperature range of 25-65 C.; stability in pH range of 6.0-10.0 and temperature 25-45 C. The protease activity was retained for 8 months when stored at ambient temperature. Ammonium sulphate precipitated enzyme was able to completely dehair animal skins and hides without chemicals like lime, sodium sulphide and calcium.

S. S. Nilegaonkar, V. P. Zambare, P. P. Kanekar, P. K. Dhakephalkar, S. S. Sarnaik, N. K. Chandrababu, Rama Rajaram, B. Ramanaiah, T. Ramasami, Y. K. Saikumari and P. Balaram (2007) A novel protease for industrial application, US Patent Patent No US20080220499A1

Link

The present invention relates to an extracellular enzyme protease obtained by growing the culture of Pseudomonas aeruginosa MCM B-327 isolated from vermiculture pit soil and deposited in MTCC, IMTECH, Chandigarh with designation MTCC 5270, in production medium of pH 7.0; containing soybean meal and tryptone as raw materials, at 30 C. for 72 h. The organism was also able to produce protease using different agricultural products/byproducts as protein sources. The partially purified non-collagenolytic, calcium independent protease with molecular weight 60 kDa has activity in pH range of 6.0-11.0 and temperature range of 25-65 C.; stability in pH range of 6.0-10.0 and temperature 25-45 C. The protease activity was retained for 8 months when stored at ambient temperature. Ammonium sulphate precipitated enzyme was able to completely dehair animal skins and hides without chemicals like lime, sodium sulphide and calcium.

S. S. Nilegaonkar, V. P. Zambare, P. P. Kanekar, P. K. Dhakephalkar, S. S. Sarnaik, N. K. Chandrababu, Rama Rajaram, B. Ramanaiah, T. Ramasami, Y. K. Saikumari and P. Balaram (2006) A novel protease for industrial application, Indian Patent Patent NO. 2471DEL2006

The present invention relates to an extracellular enzyme protease obtained by growing the culture of Pseudomonas aeruginosa MCM B-327 isolated from vermiculture pit soil and deposited in MTCC, IMTECH, Chandigarh with designation MTCC 5270, in production medium of pH 7.0; containing soybean meal and tryptone as raw materials, at 30 C. for 72 h. The organism was also able to produce protease using different agricultural products/byproducts as protein sources. The partially purified non-collagenolytic, calcium independent protease with molecular weight 60 kDa has activity in pH range of 6.0-11.0 and temperature range of 25-65 C.; stability in pH range of 6.0-10.0 and temperature 25-45 C. The protease activity was retained for 8 months when stored at ambient temperature. Ammonium sulphate precipitated enzyme was able to completely dehair animal skins and hides without chemicals like lime, sodium sulphide and calcium.

S. S. Nilegaonkar, V. P. Zambare and P. P. Kanekar (2004) Extracellular protease from Bacillus sp. BSA-26: application in dehairing of buffalo hide, Biotechnological Approaches for Sustainable Development, Allied Publishing Pvt. Ltd.

Link



...