Abstract
In order to improve the production of the milk-clotting enzyme under submerged fermentation, two statistical methods were applied to optimize the culture conditions of Bacillus amyloliquefaciens D4 using wheat bran as nutrient source. First, initial pH, agitation speed, and fermentation time were shown to have significant effects on D4 enzyme production using the Plackett–Burman experimental design. Subsequently, optimal conditions were obtained using the Box–Behnken method, which were as follows: initial pH 7.57, agitation speed 241 rpm, fermentation time 53.3 h. Under these conditions, the milk-clotting enzyme production was remarkably enhanced. The milk-clotting enzyme activity reached 1996.9 SU/mL, which was 2.92-fold higher than that of the initial culture conditions, showing that the Plackett–Burman design and Box–Behnken response surface method are effective to optimize culture conditions. The research can provide a reference for full utilization of wheat bran and the production of milk-clotting enzyme by B. amyloliquefaciens D4 under submerged fermentation.
This is a preview of subscription content, log in via an institution to check access.
References
Wim JQ (2006) Bacterial enzymes. Prokaryotes 1:777–796
Das K, Doley R, Mukherjee AK (2004) Purification and biochemical characterization of a thermostable, alkaliphilic, extra-cellular a-amylase from Bacillus subtilis DM-03, a strain isolated from the traditional fermented food of India. Appl Biochem Biotech 40:291–298
Yang HQ, Liu L, Shin HD, Chen RR, Li JH, Du GC, Chen J (2013) Structure-based engineering of histidine residues in the catalytic domain of α-amylase from Bacillus subtilis for improved protein stability and catalytic efficiency under acidic conditions. J Biotechnol 1:59–66
Annamalai N, Thavasi R, Vijayalakshmi S (2011) Extraction, purification and characterization of thermostable, alkaline tolerant α-amylase from Bacillus cereus. Indian J Microbiol 51:424–429
Christiansen T, Christensen B, Nielsen J (2002) Metabolic network analysis of Bacillus clausii on minimal and semirich medium using (13)C-labeled glucose. Metab Eng 4:159–169
Deng AH, Wu J, Zhang Y, Zhang GQ, Wen TY (2010) Purification and characterization of a surfactant-stable high-alkaline protease from Bacillus sp. B001. Bioresour Technol 18:7100–7106
Anissa H, Nahed FZ, Noomen H, Fakher F, Moncef N, Alya SK (2010) Low-cost fermentation medium for alkaline protease production by Bacillus mojavensis A21 using hulled grain of wheat and sardinella peptone. J Biosci Bioeng 3:288–294
Bindu B, Saurabh SD, Sonia A, Ritu M, Jitender S (2012) Application of thermostable xylanase of Bacillus pumilus in textile processing. Indian J Microbiol 52:222–229
Ageitos JM, Vallejo JA, Sestelo ABF (2007) Purification and characterization of a milk-clotting protease from Bacillus licheniformis strain USC13. J Appl Microbiol 103:2205–2213
Dutt K, Gupta P, Saran S (2009) Production of milk-clotting protease from Bacillus subtilis. Appl Biochem Biotech 158:761–772
Shieh CJ, Phan TL, Shih IL (2009) Milk-clotting enzymes produced by culture of Bacillus subtilis natto. Biochem Eng J 43:85–91
Ding ZY, Liu SP, Gu ZH, Zhang L, Zhang KC, Shi GY (2011) Production of milk-clotting enzyme by Bacillus subtilis B1 from wheat bran. Afr J Biotechnol 10:9370–9378
Subhosh MC, Buddolla VB, Rajasekhar R (2010) Optimization of extraction of β-endoglucanase from the fermented bran of Aspergillus niger. Indian J Microbiol 50(suppl 1):122–126
Gan BZ, Song X, He XL, Zhang WB, Li F (2009) Separation and identification of a chymosin producing bacterium from soil of yak grazing district in Tianzhu country of Gansu province. Food Sci 30(11):158–162 (in chinese)
He XL, Ren FZ, Guo HY, Zhang WB, Song X, Gan BZ (2011) Purification and properties of a milk-clotting enzyme produced by Bacillus amyloliquefaciens D4. Korean J Chem Eng 28:203–208
Acknowledgments
This work was financed by the National Key Technologies R & D Program of China in rural areas (2011AA100903) and National Key Technologies R & D Program of China (2012BAD28B07).
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Zhang, W., He, X., Liu, H. et al. Statistical Optimization of Culture Conditions for Milk-Clotting Enzyme Production by Bacillus Amyloliquefaciens Using Wheat Bran-An Agro-Industry Waste. Indian J Microbiol 53, 492–495 (2013). https://doi.org/10.1007/s12088-013-0391-2
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s12088-013-0391-2