Purification and properties of a milk-clotting enzyme produced by Bacillus amyloliquefaciens D4

He, Xiaoling; Ren, Fazheng; Guo, Huiyuan; Zhang, Weibing; Xi, Song; Bo-zhong, Gan

doi:10.1007/s11814-010-0347-8

Cited by 36 publications

(31 citation statements)

References 33 publications

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“…Milk clotting acid protease (religiosin) was reported from the latex of Ficus religiosa belonging to the serine protease group (Kumari, Sharma, & Jagannadham, 2010). Milk-clotting enzyme (YS-1) with high specificity to the substrate b-casein was purified and identified as a metalloproteinase from Bacillus subtilis (Li et al, 2012) and from Bacillus amyloliquefaciens D4 (He et al, 2011). Purification and characterisation of a milkclotting serine protease from Bacillus licheniformis strain USC13 was also reported (Ageitos, Vallejo, Sestelo, Poza, & Villa, 2007).…”

Section: Discussionmentioning

confidence: 99%

Purification and characterisation of κ-casein specific milk-clotting metalloprotease from Termitomyces clypeatus MTCC 5091

2015

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Section: Discussionmentioning

confidence: 99%

Purification and characterisation of κ-casein specific milk-clotting metalloprotease from Termitomyces clypeatus MTCC 5091

2015

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“…However, attention has been focused on the production of milk-clotting enzymes (MCEs) from microbial sources for use as rennin substitutes (Ayhan et al 2001 ; Cavalcanti et al 2004 ; Hashem 1999 ; Silveira et al 2005 ). Although there are many microorganisms that produce MCEs (Ding et al 2011 ; He et al 2011 ; Li et al 2012 ; Vishwanatha et al 2010 ), only the MCEs produced by strains of Rhizomucor miehei , Rhizomucor pusillus var. Lindt , Aspergillus oryzae and Enthothia parasitica are widely used (Birkkjaer & Jonk 1985 ; Crawford 1985 ; Thakur et al 1990a ).…”

Section: Introductionmentioning

confidence: 99%

Optimization of the production and characterization of milk clotting enzymes by Bacillus subtilis natto

Chang

Shih

2013

SpringerPlus

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Suitable medium for production of milk clotting enzyme (MCE) by Bacillus subtilis (natto) Takahashi in submerged liquid-state fermentation was screened, the nutrient factors affecting MCE production was optimized by response surface methodology. The MCE production by B. subtilis (natto) Takahashi was increased significantly by 428% in the optimal medium developed. The MCE was filtered and concentrated by ultrafiltration. The retentate after tandem filtration carried out with the combined membranes of MWCO 50kDa and 5 kDa showed two major bands between 25kDa and 30kDa on SDS-PAGE, and the MCA and MCA/PA improved significantly in comparison with those in the initial broth. The crude enzyme thus obtained showed MCA and MCA/PA ratio of 48,000 SU/g and 6,400, which are commensurate with those (MCA 26,667 SU/g and MCA/PA 6,667) of the commercial rennet. It had optimal pH and temperature at pH 6 and 60°C, and showed excellent pH and thermal stability.

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“…We recently reported several bacteria producing milk-clotting enzyme isolated from the yak grazing soil in the north-eastern Tibetan Plateau. Among these bacteria, B. amyloliquefaciens D4 possessed high rennet-producing capacity in wheat bran juice [14,15]. The objective of this work was to attempt to optimize the culture conditions to increase milk-clotting enzyme production from wheat bran by B. amyloliquefaciens D4.…”

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confidence: 99%

“…The optimum temperature and optimum pH were 65°C and 5.5, respectively [15]. Milk-clotting enzyme production by B. amyloliquefaciens has been successfully applied in the preparation of cheese (Fig.…”

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confidence: 99%

Statistical Optimization of Culture Conditions for Milk-Clotting Enzyme Production by Bacillus Amyloliquefaciens Using Wheat Bran-An Agro-Industry Waste

Zhang

Liu

et al. 2013

Indian J Microbiol

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In order to improve the production of the milkclotting 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 milkclotting 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.Keywords Optimization Á Culture Conditions Á Milkclotting enzyme Á Wheat bran Many bacteria especially several species belonging to Bacillus are known to produce variety of extracellular enzymes and they have a wide range of industrial applications [1]. Most commercial amylases are produced from a small subgroup of Bacillus species such as B. amyloliquefaciens, B. subtilis, and B. cereus [1][2][3][4].Various bacterial protease have been widely used such as B. subtilis, B. licheniformis, and B. mojavensis [5][6][7]. Xylanase from bacteria, such as B. pumilus, are being applied in textile processing [8]. Numerous bacteria belonging to Bacillus such as B. licheniformis, B. subtilis and B. subtilis natto have been suggested as promising microbial rennet producers [9][10][11][12].Wheat bran, an agro-industrial residue, a cheap source of energy has high potential in the area of fermentation for the production of enzymes. Wheat bran contains cellulose material, starch, crude protein, trace elements and other certain ingredients, which can be used as carbon and nitrogen sources to promote the growth of microorganisms and enzyme production. There are several reports describing wheat bran as potent substrate for enzyme production [12,13]. We recently reported several bacteria producing milk-clotting enzyme isolated from the yak grazing soil in the north-eastern Tibetan Plateau. Among these bacteria, B. amyloliquefaciens D4 possessed high rennet-producing capacity in wheat bran juice [14,15]. The objective of this work was to attempt to optimize the culture conditions to increase milk-clotting enzyme production from wheat bran by B. amyloliquefaciens D4.The producer strain was B. amyloliquefaciens D4(CGMCC 3290), which was deposited in the China General Microbiological Culture Collection Center (Beijing, China). The strain was propagated at 37°C on lysogeny broth (LB) agar slants (1.0 % (w/v) peptone, 1.0 % (w/v) beef extract, 0.5 % (w/v) NaCl, and 2.0 % (...

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Purification and properties of a milk-clotting enzyme produced by Bacillus amyloliquefaciens D4

Cited by 36 publications

References 33 publications

Purification and characterisation of κ-casein specific milk-clotting metalloprotease from Termitomyces clypeatus MTCC 5091

Purification and characterisation of κ-casein specific milk-clotting metalloprotease from Termitomyces clypeatus MTCC 5091

Optimization of the production and characterization of milk clotting enzymes by Bacillus subtilis natto

Statistical Optimization of Culture Conditions for Milk-Clotting Enzyme Production by Bacillus Amyloliquefaciens Using Wheat Bran-An Agro-Industry Waste

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