长鳍金枪鱼多肽的酶解制备工艺研究
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摘要
通过酶解法制备具有对H_2O_2诱导的张氏肝细胞保护作用的长鳍金枪鱼多肽。以长鳍金枪鱼下脚料为原料,采用胰蛋白酶进行酶解,以张氏肝细胞的相对增殖率为指标,通过单因素试验和响应面优化试验确定该酶的最佳酶解条件, HE染色观察张氏肝细胞形态变化。结果表明,最佳酶解条件为:料酶解pH 7.28,酶解温度45.29℃,时间5.05 h,所得酶解产物对张氏肝细胞的相对增殖率为53.1%。HE染色显示长鳍金枪鱼多肽对H_2O_2诱导的张氏肝细胞起一定保护作用。该工艺研究为长鳍金枪鱼多肽的分离纯化及开发护肝功能食品提供试验依据。
The T. alalunga polypeptide with protection about H_2 O_2 induced liver cell was prepared by enzymatic hydrolysis. The T. alalunga scraps were used as the raw materials and were prepared for trypsin digestion. The optimal enzymolysis condition was determined by single factor experiments and experimental tests which were screened using relative growth rate as the indicator. Hematoxylin-eosin staining was observed the morphological changes of Chang liver. The results showed that the optimal enzymatic hydrolysis condition was as follows: p H 7.28, temperature maintained at 45.29 ℃ for 5.05 h. The enzymatic hydrolysis product of Chang liver cells relative proliferation rate was 53.1%. Hematoxylin-eosin staining showed that the T.alalunga polypeptide had a protective effect on H_2 O_2 induced Chang liver cells. An experimental basis for the separation and purification of T. alalunga polypeptide and the development of liver function healthy food was provided.
The T. alalunga polypeptide with protection about H_2 O_2 induced liver cell was prepared by enzymatic hydrolysis. The T. alalunga scraps were used as the raw materials and were prepared for trypsin digestion. The optimal enzymolysis condition was determined by single factor experiments and experimental tests which were screened using relative growth rate as the indicator. Hematoxylin-eosin staining was observed the morphological changes of Chang liver. The results showed that the optimal enzymatic hydrolysis condition was as follows: p H 7.28, temperature maintained at 45.29 ℃ for 5.05 h. The enzymatic hydrolysis product of Chang liver cells relative proliferation rate was 53.1%. Hematoxylin-eosin staining showed that the T.alalunga polypeptide had a protective effect on H_2 O_2 induced Chang liver cells. An experimental basis for the separation and purification of T. alalunga polypeptide and the development of liver function healthy food was provided.
引文
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[6] LOMINO J V, NAEGELI A, ORWENYO J, et al. A twostep enzymatic glycosylation of polypeptides with complex N-glycans[J]. Bioorganic&Medicinal Chemistry, 2013,21(8):2262.
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[8]顾斌洲,王加斌,滕芳芳,等.鳕鱼皮制备胶原蛋白肽的工艺优化[J].浙江海洋学院学报(自然科学版), 2015, 34(5):452-456.
[9] WAGLAY A, KARBOUNE S. A novel enzymatic approach based on the use of multi-enzymatic systems for the recovery of enriched protein extracts from potato pulp[J]. Food Chemistry,2017, 220:313-323.
[10] TRUSEK-HOLOWNIA A, LECH M, NOWORYTA A. Protein enzymatic hydrolysis integrated with ultrafiltration:Thermolysin application in obtaining peptides[J]. Chemical Engineering Journal, 2016, 305:61-68.
[11] AZEEM A K, DILIP C, PRASANTH S S, et al. Antiinflammatory activity of the glandular extracts of Thunnus alalunga[J]. Asian Pacific Journal of Tropical Medicine, 2010,3(10):794-796.
[2]向泽敏,葛俊驿,邱晓挺,等.鲭科鱼类活性肽的酶法制备及其生理功能研究进展[J].生物学杂志, 2017, 34(1):65-69.
[3] YANG S, GAO Q, XING S, et al. Neuroprotective effects of Buyang Huanwu decoction against hydrogen peroxide induced oxidative injury in Schwann cells[J]. Journal of Ethnopharmacology, 2011, 137(3):1095-1101.
[4] LIAO H, JIANG L, WEI C, et al. Applied orthogonal experiment design for the optimum extraction conditions of high concentration selenium from Maifanite[J]. Detection,2017, 5(1):1-4.
[5]罗李王,杨最素,张亚茹,等.酶解青蛤制备ACE抑制肽的工艺优化[J].食品工业, 2016, 37(6):56-59.
[6] LOMINO J V, NAEGELI A, ORWENYO J, et al. A twostep enzymatic glycosylation of polypeptides with complex N-glycans[J]. Bioorganic&Medicinal Chemistry, 2013,21(8):2262.
[7] NONGONIERMA A B, FITZGERALD R J. Unlocking the biological potential of proteins from edible insects through enzymatic hydrolysis:A review[J]. Innovative Food Science&Emerging Technologies, 2017:43.
[8]顾斌洲,王加斌,滕芳芳,等.鳕鱼皮制备胶原蛋白肽的工艺优化[J].浙江海洋学院学报(自然科学版), 2015, 34(5):452-456.
[9] WAGLAY A, KARBOUNE S. A novel enzymatic approach based on the use of multi-enzymatic systems for the recovery of enriched protein extracts from potato pulp[J]. Food Chemistry,2017, 220:313-323.
[10] TRUSEK-HOLOWNIA A, LECH M, NOWORYTA A. Protein enzymatic hydrolysis integrated with ultrafiltration:Thermolysin application in obtaining peptides[J]. Chemical Engineering Journal, 2016, 305:61-68.
[11] AZEEM A K, DILIP C, PRASANTH S S, et al. Antiinflammatory activity of the glandular extracts of Thunnus alalunga[J]. Asian Pacific Journal of Tropical Medicine, 2010,3(10):794-796.