亮斑扁角水虻幼虫酶解制备蛋白胨工艺优化及蛋白胨性质

Peptone prepared from Hermetia illucens larvae: optimization of enzymolysis conditions and characteristics

摘要:

【背景】用于餐厨垃圾等有机废弃物处理的亮斑扁角水虻(Hermetia illucens, HI)含有丰富的氨基酸和营养物质，是一种优质、经济的蛋白来源。【目的】利用亮斑扁角水虻幼虫(Hermetia illucens larvae, HIL)制备蛋白胨用于细菌培养，为亮斑扁角水虻的应用方式提供新的思路。【方法】以亮斑扁角水虻幼虫为原料，利用单因素试验确定最佳酶解条件，对比分析HIL蛋白胨和市售胰蛋白胨的基本性质和功能性状，并进行细菌培养、生物化学试验，利用两种蛋白胨分别培养大肠杆菌(Escherichia coli) ATCC 25922模式细菌并通过生长动力学分析评价应用效果。【结果】制备HIL蛋白胨的最佳酶解工艺为按1.3% (质量分数)添加复合酶(胰酶:碱性蛋白酶质量比为1:1)，在pH 7.0、54 ℃条件下反应4 h，此时HIL的水解度为(19.34±0.15)%。HIL蛋白胨和市售胰蛋白胨的功能性状和生物化学试验差异不显著(p>0.05)。大肠杆菌ATCC 25922在HIL蛋白胨培养基中生长的X_max和λ分别为6.44和2.45，在市售胰蛋白胨生长的X_max和λ分别为6.14和3.19，差异均不显著(p>0.05)。【结论】在培养大肠杆菌时，HIL蛋白胨可以替代胰蛋白胨作为微生物培养基的成分。

Abstract:

[Background] Hermetia illucens (HI) used for the treatment of organic wastes such as kitchen waste is rich in amino acids and nutrients, serving as a high-quality and economical source of protein. [Objective] To prepare peptone with H. illucens larvae (HIL) for bacterial culture and provide a new idea for the application of HI. [Methods] Peptone was prepared from HIL by enzymolysis. The optimal enzymolysis conditions were determined by single factor tests, and the biochemical and functional properties of HIL-derived peptone and commercial tryptone were compared. Escherichia coli ATCC25922 was cultured with two peptone products and the growth kinetics was compared. [Results] The optimal conditions for preparing HIL-derived peptone were enzymolysis with the enzyme complex (trypsin:alkaline protease=1:1) added at 1.3% (mass fraction ) and at pH 7.0 and 54 ℃ for 4 h, under which the hydrolysis degree of defatted HIL was (19.34±0.15)%. The functional and biochemical properties showed no significant differences between HIL-derived peptone and commercial tryptone (P>0.05). The X_max,and λ of E. coli ATCC 25922 grown in the medium supplemented with HIL-derived peptone were 6.44 and 2.45, respectively, and those of E. coli ATCC 25922 grown in the medium supplemented with commercial tryptone were 6.14 and 3.19, respectively. The X_max,and λ showed no significant differences between HIL-derived peptone and commercial tryptone (p>0.05).[Conclusion] HIL-derived peptone can replace commercial tryptone as a component of the medium for culturing E. coli.