为充分利用螺旋藻渣副产物,获得较佳的抗菌活性发酵产物,该文以团队筛选的枯草芽孢杆菌(Bacillus subtilis YA215)为发酵剂,对螺旋藻渣进行液态发酵。以发酵上清液中性蛋白酶活、多肽含量及对金黄色葡萄球菌(Staphylococcus aureus)抑菌率为指标,通过单因素实验、Plackett-Burman实验和正交试验,优化发酵工艺条件。结果表明,发酵初始溶液中,螺旋藻渣添加量15 g/L、果糖添加量10 g/L、NaCl添加量10 g/L、YA215接种量75 mL/L(发酵剂菌落含量为107~108CFU/mL)、pH 7.0,37 ℃摇床培养48 h(转速200 r/min),其发酵上清液中性蛋白酶活达到147.4 U/mL,多肽含量为16.8 mg/mL,对金黄色葡萄球菌抑菌率为64.6%,产生的抑菌圈直径大小为15.3 mm,且抑菌率与多肽含量正相关。螺旋藻渣可通过枯草芽孢杆菌YA215发酵获得良好抗菌性,提升其作为饲料配料的价值。
The purpose of this study was to make full use of the by-products of spirulina residue to obtain higher antimicrobial activity fermented products. In this study, Bacillus subtilis YA215 was selected as the seeding culture for spirulina residue fermentation. The neutral protease activity, polypeptide content and antimicrobial activity (against Staphylococcus aureus) of the supernatant were analyzed and used as indexes. Based on the indexes, the fermentation conditions were optimized using the single factor, Plackett-Burman, and orthogonal experiment methods. The optimized condition as follows: 1.5% (w/w) of spirulina residue, 1% (w/w) of fructose, 1% (w/w) of NaCl, and 7.5% (v/v) of YA215 starter culture (107-108 CFU/mL), pH 7.0 and under 37 ℃ for 48 h with agitation of 200 r/min. The results showed that the neutral protease activity of the fermentation supernatant reached 147.4 U/mL. Moreover, the polypeptide reached 16.8 mg/mL and antimicrobial rate was 64.6%. Meanwhile the diameter of the produced bacteriostatic circle was 15.3 mm. In addition, the antimicrobial activity was positively related to the polypeptide content. In summary, the spirulina residue can be fermented using Bacillus subtilis YA215 to obtain high antibacterial property. This product has a high potential value in feed application.
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