枯草芽孢杆菌等四菌组合固态发酵工艺参数的优化

Optimization of process parameters for solid-state fermentation by four-strain combination including Bacillus subtilis

摘要

为优化枯草芽孢杆菌(Bacillus subtilis,BS)、热带假丝酵母(Candida tropicalis,CT)、黑曲霉(Aspergillus niger,AN)、宇佐美曲霉(Aspergillus usamii,AU)四菌组合固态发酵工艺参数,开发高营养复合益生菌发酵饲料,采用平板刺激圈实验验证菌株相容性,以粗蛋白含量为核心指标,通过单因素实验筛选关键影响因子范围,结合正交试验优化工艺参数.结果显示:四菌最佳接种比例为枯草芽孢杆菌(BS)∶热带假丝酵母(CT)∶黑曲霉(AN)∶宇佐美曲霉(AU)=4∶2∶3∶4;该条件下,发酵产物粗蛋白含量达22.384 mg/g,较对照组提高54.6%,且四菌组合发酵效果显著优于单菌及两菌组合发酵效果.按此四菌组合配比进行固态发酵,最优工艺参数为接种量20%、麸皮含水量60%、发酵温度32 ℃、发酵时间60 h,发酵产物粗蛋白含量为23.795 mg/g,较对照组提高64.3%.研究表明,四菌间存在协同互补效应,优化后的工艺参数可为复合益生菌发酵饲料的工业化生产提供技术支撑.

Abstract

To optimize the solid-state fermentation process parameters of a four-strain microbial consortium consisting of Bacillus subtilis, Candida tropicalis, Aspergillus niger and Aspergillus usamii and to develop high-nutrition compound probiotic fermented feed, the inter-strain compatibility was verified using a plate stimulation circle assay. Taking crude protein content as the core evaluation index, single-factor experiments were conducted to screen the range of key influencing factors, and the process parameters were optimized via orthogonal experiments. The results demonstrated that the optimal inoculation ratio of the four strains was Bacillus subtilis∶Candida tropicalis∶Aspergillus niger∶Aspergillus usamii=4∶2∶3∶4. Under this condition, the crude protein content of the fermented product reached 22.384 mg/g, which was 54.6% higher than that of the control group. Meanwhile, the fermentation performance of the four-strain mixed fermentation was significantly superior to that of single-strain and two-strain combined fermentation. According to this ratio for solid-state fermentation, the optimal process parameters were obtained with inoculum size of 20%, wheat bran moisture content of 60%, fermentation temperature of 32 ℃, and fermentation duration of 60 h. Under these conditions, the crude protein content of the fermented product reached 23.795 mg/g, which was 64.3% higher than that of the control group. This study indicates that there is a synergistic and complementary effect among the four strains and the optimized process parameters can provide technical support for the industrial production of compound probiotic fermented feed.

关键词

四菌组合 / 固态发酵 / 工艺参数优化 / 粗蛋白 / 发酵饲料

Key words

four-strain combination / solid-state fermentation / process parameter optimization / crude protein / fermented feed

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