ZENG Qing-ranet al: Study on Various Enzymatic Hydrolysis of Chicken Skin for Collagen Peptides Research Article PAGE : ( 3 313~20 100); Tris Tricine

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1 曾庆冉等 : 多种酶复合制备鸡皮胶原蛋白短肽曾庆冉 1 张慜 *1 陈世豪 2 ( ; ) : 以鸡皮为原料采用酶法提取胶原蛋白肽研究了酶的种类加酶量酶解时间酶解温度以及 ph 值对水解度和多肽质量浓度的影响采用正交实验对酶解条件进行了优化结果表明当采用木瓜蛋白酶 + 胰蛋白酶 + 风味蛋白酶复合水解加酶量为 0.9% 酶解时间 3 h 酶解温度 45 及 ph 为 7 时为酶解最佳条件 : 鸡皮 ; 胶原蛋白肽 ; 水解 ; 优化 :Q 516 :A : (2013) Study on Various Enzymatic Hydrolysis of Chicken Skin for Collagen Peptides ZENG Qing-ran 1 ZHANG Min *1 CHEN Shi-hao 2 (1. School of Food Science and TechnologyJiangnan UniversityWuxi China;2. Guangdong Jiahao Food Ltd. CoZhongshan China) Abstract: The collagen peptides from chicken skin has been extracted by using enzyme method. In additionthe effects of enzyme typesenzyme concentrationhydrolysis timehydrolysis temperature and ph on peptide hydrolysis were studiedand the extraction conditions were optimized by orthogonal experiment. The results shows that the optimum enzyme was papain add trypsin add flavorzyme and the optimum concentration of enzyme was 0.9%the time of enzyme hydrolysis was 3 hoursthe temperature of enzyme hydrolysis was 45 and ph was 7. Keywords: chicken skincollagen peptideshydrolysisoptimization 2~6 [1] [2] : : 863 (2011AA100802) * : (1962 ) min@jiangnan.edu.cn 貋貙貪

2 ZENG Qing-ranet al: Study on Various Enzymatic Hydrolysis of Chicken Skin for Collagen Peptides Research Article PAGE : ( 3 313~20 100); Tris Tricine TEMED : 1.3 Siriporn Damrongsakkul ph FE20: - ( [3] ) ; PL203: - [4] ( ) ;CJJ78-1 : ;SIGMA2-16PK : ;HH-S : [5] ;UV-9100 : [6] ;U410 :New BRUNSWICK SCIENTIFIC CO.; : ;DYY-8C : ; 原料预处理 10 g/dl [7] : 鸡皮胶原蛋白肽制备工艺 Table 1 Material's mainly composition min 1.2 /% /% /% /% /% 0.45 SDS NN'- NaCl ( 1 10)4 12 h ph 10 g/dl 4 24 h r/min 10 min [8-9] 分析测定方法 [10] 1) : 2) : 5 ml 10% 5 ml 10 min r/min 20 min (1 cm 1 cm) -20 [11] 3) : 5 ml 6 mol/l 10 ml 110 Flavourzyme PROTAMEX 24 h 50 ml Novozym 37071: SDS- 1 ml 5 ml 貋貚財 Journal of Food Science and Biotechnology Vol.32 No

1 1 [7] : 1 1.4.2 鸡皮胶原蛋白肽制备工艺 1 1 10 Table 1 Material's mainly composition 85 10 min 1.2 /% /% 50.58 /% 37.62 /% 11.14 /% 0.

3 曾庆冉等 : 多种酶复合制备鸡皮胶原蛋白短肽 40~50 1 ml ph 2.2 [12] 4) : Tricine-SDS-PAGE [13-15] V;1 h 100 V (0.5% 30% ddh 2 O 100 ml) 30 min G min 1 2 Fig. 1 Effects of enzymatic types on peptide hydrolysis Table 2 Polyacrylamide gel formula 2.2 ph 49.5%T 3%C ml ml 49.5%T 5%C 2 ml 2 ml ml ml g ddh 2O ml ml 10%AP 80 μl 20 μl 20 μl TEMED 10 μl 5 μl 5 μl 2.1 Flavourzyme PROTAMEX Novozym ( ph ) 2 ( + + ) 3 Fig. 2 Effects of ph on peptide hydrolysis ( + + ) (Novozym ) 5( + + ) ph 7 1% % 50 3 h ( h ) [16] % 50 ph 5~8.5 3 h ph 2 2 ph 5~8.5 ph ph 7 ; ph 7.5 ph 7~7.5 ph 7~ 貋貚貢

926 ml 1.344 ml 10%AP 80 μl 20 μl 20 μl TEMED 10 μl 5 μl 5 μl 2.1 Flavourzyme PROTAMEX Novozym 37071 1( + + 2 ph ) 2 ( + + ) 3 Fig. 2 Effects of ph on peptide hydrolysis ( + + ) 4 2.

4 ZENG Qing-ranet al: Study on Various Enzymatic Hydrolysis of Chicken Skin for Collagen Peptides Research Article Fig. 3 3 h ph 7 0.8% 3 h 3 Effects of hydrolysis time on peptide hydrolysis ph h % Fig. 5 Effects of hydrolysis temperature on peptide 0.8% hydrolysis 2.6 ph 3 3 > > >ph ph h 0.9% 0.8% 5 35~ mg/ml 多肽氨基酸含量及组成分析 % % 4 4 Fig. 4 Effects of enzyme concentrations on peptide hydrolysis 貋貚貣 Journal of Food Science and Biotechnology Vol.32 No

5 Effects of hydrolysis temperature on peptide 0.8% hydrolysis 2.6 ph 3 3 > > >ph ph 7 45 3 h 0.9% 0.8% 5 35~50 50 50 50 1.32 mg/ml 2.7 2.

5 曾庆冉等 : 多种酶复合制备鸡皮胶原蛋白短肽 [17] Table 3 3 Table 4 Amino acid composition of collagen polypeptides Design and results of orthogonal experiment for optimization of hydrolysis conditions 多肽相对分子质量测定 SDS-PAGE 6 M : Tricine-SDS-PAGE : 5 856~ ~ ph 7 0.9% 45 3 h 2 000~ ph / /h /% / (mg/ml) K K K R SDS-PAGE Fig. 6 SDS -PAGE electrophoretogram of collagen polypeptide 3 /% /% : [ 1 ]. [J]. 2006(2): LI Er-fengHE Xiao-weiLUO Zhi-gang. Application of collagen to food industry[j]. China Dairy Industry2006(2): (in Chinese) [ 2 ]. [J]. 2005(9): ZHAO LiSU WeiHU Huo -genet al. Research progress of collagen peptides [J]. Food Science2005 (9): (in Chinese) [ 3 ] Siriporn DamrongsakkulKongpob RatanathammapanKittinan Komolpiset al. Enzymatic hydrolysis of rawhide using papain and neutrase[j]. Journal of Industrial and Engineering Chemistry2008(14): [ 4 ]. [J]. 2010(5): LI Xing-wuZHANG Li-li. Preparation of collagen peptide from pig by-products by enzymatic way [J]. Meat Research 貋貚貤

37 1 6.5 45 2.5 0.7 0.78 2 6.5 50 3 0.8 0.68 3 6.5 55 3.5 0.9 0.73 4 7 45 3 0.9 1.27 5 7 50 3.5 0.7 0.71 6 7 55 2.5 0.8 0.68 7 7.5 45 3.5 0.8 0.82 8 7.5 50 2.5 0.9 0.68 9 7.5 55 3 0.7 0.68 K 1 0.

6 ZENG Qing-ranet al: Study on Various Enzymatic Hydrolysis of Chicken Skin for Collagen Peptides Research Article (5):83-87.(in Chinese) [ 5 ] M C Gomez-GuillenB Gimenez. Functional and bioactive properties of collagen and gelatin from alternative sources:a review [J]. Food Hydrocolloids2011(25): [ 6 ]. [J]. 2007(4): JIA Dong-yingZHAO Hong-ming. Enzymatic extraction of protein from the head and bone of silver carp by papain [J]. Food Science And Technology2007(4): (in Chinese) [ 7 ] Prabjeet SinghSoottawat Benjakul. Isolation and characterisation of collagen extracted from the skin of striped catfish (Pangasianodon hypophthalmus)[j]. Food Chemistry2011(124): [ 8 ] Krzysztof SurowkaM Fik. Studies on the recovery of proteinaceous substances from chicken heads: -application of pepsin to the production of protein hydrolysate[j]. J Sci Food Agric1994(65): [ 9 ]. [J]. 2011(4): SONG QinYAN Jun. Study on extracion technology of tilapia fish scale collagen oligopeptide [J]. Food Research And Development2011(4): (in Chinese) [10]. [M]. : [11]. [J]. 2005(7): LU WeiREN Guo-puSONG Jun-mei. Determination of content of peptides in protein hydrolysates[j]. Food Science2005(7): (in Chinese) [12] Jinhan ShonJi-Hyun Eo. Effect of processing conditions on functional properties of collagen powder from skate (Raja kenojei) skins[j]. Food Sci Biotechnol2011(1): [13]. 1kDa Tricine-SDS-PAGE [J]. 2004(1): CAO Zuo-wu. Tricine-SDS-PAGE for the effective separation of 1KDa peptides[j]. China Biotechnology2004(1): (in Chinese) [14]. Tricine-SDS-PAGE [J]. 2003(2): WANG XuHE Bing-fangLI Shuanget al. Tricine-SDS-PAGE electrophoresis analysis of low molecular peptides [J]. Journal of Nanjing University of Technology2003(2):79-81.(in Chinese) [15]. SDS-PAGE [J]. 2001(1): SHI Ji -hongzhao Yong -tongzhang Ying -qiet al. Study on analysing methods of low molecular peptides with SDS - polyacrylamide Gel electrophoresis[j]. Biological Project Progress2001(1): (in Chinese) [16]. [J]. 2010(7): SHAO WeiDENG Yuan-fangCHEN Shu. Production of collagen peptide from bullhead skin by peptide hydrolase [J]. China Brewing2010(7): (in Chinese) [17]. [J]. 2009(4): LIANG Yong -junsun Xiang -jun. Study on the preparation and physicochemical characteristics of bioactive peptides from tilapia skin collagen[j]. Journal of Anhui Agri Sci2009(4): (in Chinese) 貋貚貥 Journal of Food Science and Biotechnology Vol.32 No

Enzymatic extraction of protein from the head and bone of silver carp by papain [J]. Food Science And Technology2007(4):243-246.(in Chinese) [ 7 ] Prabjeet SinghSoottawat Benjakul.

结果与讨论

结果与讨论倪卓杜学晓王帅邢锋以脲醛树脂为囊壁双酚型环氧树脂为囊芯采用原位聚合法合成环氧树脂微胶囊根据化学反应动力学得到脲醛树脂预聚体及微胶囊囊壁合成反应过程中的速率方程测定不同温度下甲醛浓度随时间变化的曲线确定不