以罗非鱼鱼皮为原料,利用枯草芽孢杆菌固态发酵(solid-state fermentation,SSF)制备胶原蛋白肽。以胶原酶酶活力为评价指标,优化产酶发酵条件,并对发酵产物进行氨基酸组成分析、超滤分离及体外抗氧化活性的研究。结果表明,最佳发酵条件为培养基初始pH值为7,种子液菌龄为14 h,加水量为10 mL,发酵温度为35 ℃,发酵时间为24 h,在此条件下胶原酶酶活力为90.05 U/mL,水解度高达41.73%。经超滤分离得到3种不同分子质量(molecular weight,MW)范围的胶原蛋白肽组分:TSCP-a(MW>10 kDa)、TSCP-b(MW 5 k~10 kDa)、TSCP-c(MW<5 kDa)。其中TSCP-c的抗氧化活性最高,其羟自由基(·OH)、超氧阴离子自由基(·$O_2^-$)和1,1-二苯基-2-三硝基苯肼(1-diphenyl-2-picrylhydrazy,DPPH)自由基清除率的半抑制浓度(half inhhibitory concentration,IC50)分别为3.83、4.19和2.37 mg/mL。该新型制备工艺简单高效,为罗非鱼皮的深入开发利用和胶原蛋白肽的生物技术生产途径提供了理论指导。
Collagen peptides were prepared from tilapia skin by solid-state fermentation with Bacillus subtilis. The activity of collagenase was selected as evaluation index to optimize the fermentation conditions and the amino acid composition, ultrafiltration, the in vitro antioxidant activity of tilapia skin hydrolysates was performed. The results showed that the optimum fermentation conditions were initial pH 7, cell age 14 h, water addition 10 mL, 35 ℃ and fermentation time 24 h. Under such conditions, the activity of collagenase was 89.94 U/mL and the hydrolysis degree of the tilapia skin collagen could reach 41.73%. The hydrolysis collagen was separated by ultrafiltration to prepare three components of polypeptide with different molecular weight range, including TSCP-a (>10 kDa), TSCP-b (5-10 kDa), TSCP-c (<5 kDa). Among which, TSCP-c had the highest antioxidant activity, the IC50 for the hydroxyl radical scavenging rate, the superoxide anion radical scavenging rate and the DPPH radical scavenging rate were 3.83, 4.19 and 2.37 mg/mL, respectively. The novel preparation process is simple and efficient, providing a theoretical guidance for further development and utilization of tilapia skin and biotech production of collagen peptides.
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