响应面法优化地衣芽孢杆菌发酵鸡骨粉产抗氧化肽的工艺

Optimisation of antioxidant peptide production from chicken bone meal fermented with Bacillus licheniformis by response surface methodology

摘要:

为了鸡骨资源的高效利用及新型抗氧化剂的开发，该文以地衣芽孢杆菌为发酵菌株，发酵鸡骨生产抗氧化肽。首先研究添加葡萄糖对地衣芽孢杆菌生长的影响，然后通过单因素试验和响应面试验优化发酵条件，同时对鸡骨发酵液进行Tricine-SDS-PAGE鉴定，最后对发酵液中的鸡骨肽进行有机膜分离，并测定不同组分抗氧化活性。结果表明，添加葡萄糖抑制地衣芽孢杆菌生长，优化最佳发酵条件为：添加鸡骨粉的质量分数4.2%，发酵pH值6.0，菌的体积分数4.0%，装液量50 mL/250 mL，发酵时间24 h。在此条件下，发酵液水解度为56.29%，DPPH·清除率为57.11%。Tricine-SDS-PAGE证实鸡骨粉已被水解。有机膜分离得到的抗氧化肽组分中，组分Ⅴ（分子量3 kDa~5 kDa的肽段）抗氧化活性最好，DPPH·清除率为59.08%，超氧阴离子清除率在19%以上，ABTS·清除率为85%。该文为提高鸡骨的综合利用价值奠定基础，拓宽了功能性产品的研发范围。

Abstract:

For the efficient use of chicken bone resources and the development of novel antioxidants, antioxidant peptides were fermented from chicken bones using Bacillus licheniformis. Firstly, the effect of glucose addition on the growth of Bacillus licheniformis was studied, and then the fermentation conditions were optimized by single factor test and response surface test. At the same time, Tricine-SDS-PAGE analysis was also performed on the chicken bone fermentation broth. Finally, the chicken bone peptides with varying molecular weights were separated by organic membrane, and their antioxidant activities were determined. The results demonstrated that the addition of glucose inhibited the growth of Bacillus licheniformis. The optimal fermentation conditions were determined as follows: the addition of chicken bone meal was 4.2%, the fermentation pH was 6.0, the bacterial inoculum was 4.0%, the liquid loading was 50 mL/250 mL, and the fermentation time was 24 h. Under these conditions, the hydrolysis degree of fermentation solution was 56.29% and the DPPH· clearance rate was 57.11%. Tricine-SDS-PAGE gel confirmed that the chicken bone meal had been hydrolyzed. Among the antioxidant peptide fractions obtained by organic membrane separation, fraction V (with a molecular weight of 3 kDa~5 kDa peptide) exhibited the highest antioxidant activity, in which the scavenging of DPPH, superoxide anion and ABTS were 59.08%, over 19% and 85% respectively. This study laid the foundation for improving the comprehensive utilization value of chicken bones and broadened the scope of research and development of functional products.