Bacillus cereus 胞外胶原蛋白酶水解牛骨胶原蛋白的动力学

食品科学 ›› 2012, Vol. 33 ›› Issue (21): 192-195.

Bacillus cereus 胞外胶原蛋白酶水解牛骨胶原蛋白的动力学

刘丽莉，杨协力，任广跃，段续

河南科技大学食品与生物工程学院

收稿日期:2011-09-20 修回日期:2012-09-20 出版日期:2012-11-15 发布日期:2012-11-09
通讯作者: 刘丽莉 E-mail:yangliuyilang@126.com
基金资助:
“十一五”国家科技支撑计划项目

Hydrolysis Kinetics of Bovine Bone Collagen by Bacillus cereus Extracellular Collagenase

Received:2011-09-20 Revised:2012-09-20 Online:2012-11-15 Published:2012-11-09
Contact: LIU Li-li E-mail:yangliuyilang@126.com

摘要/Abstract

摘要： 针对从细菌蜡样芽孢杆菌(Bacillus cereus)MBL13中提取得到的胶原蛋白酶酶解牛骨胶原蛋白的动力学性质进行研究。考察底物质量浓度、时间、温度、酶质量浓度、pH值5个因素对酶解水解度的影响，优化酶解的条件；并对酶解过程的动力学进行分析。结果表明：最佳的酶解工艺参数为底物质量浓度30g/L、时间6h、温度45℃、初始酶液质量浓度为0.35g/100mL、pH8.0。在此基础上推导出酶解动力学方程为：V =9.3633[S]/(114.785+[S])。并在波长230nm和570nm处对酶解过程动态变化进行测定，表明胶原多肽含量不断增加的动态变化。

关键词: 胶原蛋白酶, 牛骨胶原蛋白, 胶原多肽, 酶解, 动力学

Abstract: In order to explore the hydrolysis kinetics of bovine bone collagen by collagenase isolated from Bacillus cereus MBL13 and the optimal hydrolysis conditions, the effects of substrate concentration, hydrolysis temperature, enzyme concentration, hydrolysis pH and hydrolysis time on the degree of hydrolysis (DH) were analyzed. The optimal conditions for the hydrolysis of bovine bone collagen were bone collagen concentration of 30 g/L, hydrolysis time of 6 h, hydrolysis temperature of 45 ℃, initial enzyme concentration of 0.35 g/100 mL and hydrolysis pH of 8.0. Based on the experimental data, a kinetic model equation was obtained as V = 9.3633·[S]/(114.785+[S]). Enzymatic hydrolysis process was analyzed at the wavelength of 230 nm and 570 nm.

Key words: collagenase, bovine bone collagen, collagen polypeptide, enzymatic hydrolysis, kinetics

中图分类号:

TS251

刘丽莉，杨协力，任广跃，段?续 . Bacillus cereus 胞外胶原蛋白酶水解牛骨胶原蛋白的动力学[J]. 食品科学, 2012, 33(21): 192-195.

参考文献

[1] David Olsen, Chunlin Yang, Michael Bodo, et al. Recombinant collagen and gelatin for drug delivery[J]. Advanced Drug Delivery Reviews, 2003, 55(12): 1547–1567.
[2] K. Gelse, E. P?schl, T. Aigner. Collagens—structure, function, and biosynthesis[J]. Advanced Drug Delivery Reviews, 2003, 55(12): 1531–1546.
[3] S. Morimura, H., Nagata, Y., Uemura, A. Fahmi, I., Shigematsu, K. Kida. Development of an effective p rocess for utilization of collagen from livestock and fish waste[J].Process Biochemistry, 2002, 7 (3):1403–1412.
[4] Chambers J. Rasmussen P.B. Enzymatic treatment of bones[J]. J.Sci.Food Agric, 1988, 42:87–94.
[5] Krzysztof Surowka, Miroslaw Fik. Studies on the Recovery of Proteinaceous Substances from Chicken Heads[J]. J.Food.Sci.Technol, 1992, 27:9–12.
[6] Michel Linder, P Rozan, et al. Protein recovery from Veal Bones by Enzymatic Hydrolysis[J]. J.Food Sci, 1995, 60 (5):949–958.
[7] Michel Linder, et al. Functional properties of veal bone hydrolysates[J]. J.Food Sci.1996, 61(7):12–720.
[8] Michel Linder, P. Rozan, et al. Nutritional Value of Veal Bone Hydrolysate[J]. J. Food Sci, 1997, 62(1):183–189.
[9] 王朝旭, 赵丹, 王小雪, 等. 酶法水解骨蛋白最佳条件的研究[J]. 食品科学, 2001, 22(2):48–49.
[10] 孙骞,王峥峥,罗永康,等. 猪骨蛋白生物活性肽酶法制备工艺的研究[J]. 肉类研究, 2008(5):37–41.
[11] K. Watanabe. Collagenolytic proteases from bacteria[J]. Appl Microbiol Biotechnol , 2004, 63: 520–526.
[12] Park PJ, Lee SH, Byun GH, et al. Purification and characterization of a collagenase mackerel, Scomber japonicus[J]. Journal od Biochemistry and Molecular Biology, 2002, 35(6): 576-582.
[13] Marilia M. Horn, Virginia C., Amaro Martins, et al. Interaction of anionic collagen with chitosan: Effect on thermal and morphological characteristics[J]. Carbohydrate Polymers, 2009, 77(2): 239-243.
[14] Miladinov V. D., Gennadios A., Hanna M. A. Gelatin manufacturing process and product[M]. US, 0142368A1. 2002.
[15] 刘丽莉，马美湖，杨协力. 牛骨I型胶原蛋白提取及结构表征[J]. 食品科学, 2010, 31(2):87–91.
[16] Nomura Y, Sakai H, Ishii Y, et al. Preparation and some properties of type I collagen from fish scales[J]. Biosci . Biotech. Biochem, 1996, 60(12) :2092–2094.
[17] Meara G M O, Munro P A. Effects of reaction variables on the hydrolysis of lean beef tissue by alcalase[J]. Meat Science, 1984, 11:227–238.
[18] Bradford MM. A rapid and sensitive method for the quantitation of microgram quantities of protein using the principle of protein-dye binding[J]. Anal Biochem, 1976, 72:248–254.
[19] T.Sonobe, N Worasuwannarak. Kinetic analyses of biomass pyrolysis using the distributed activation energy model[J]. Fuel, 2008, 87(3):414–421.