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利用实时 PCR 和变性梯度凝胶电泳(DGGE)揭示农业工业废物生产聚-γ-谷氨酸固态发酵过程中细菌群落的动态变化。

Dynamics of bacterial communities during solid-state fermentation using agro-industrial wastes to produce poly-γ-glutamic acid, revealed by real-time PCR and denaturing gradient gel electrophoresis (DGGE).

机构信息

Jiangsu Provincial Key Lab for Organic Solid Waste Utilization, Nanjing Agricultural University, Nanjing 210095, China.

出版信息

Appl Microbiol Biotechnol. 2011 Nov;92(4):717-25. doi: 10.1007/s00253-011-3375-3. Epub 2011 Jun 14.

DOI:10.1007/s00253-011-3375-3
PMID:21670980
Abstract

The dynamics of bacterial communities play an important role in solid-state fermentation (SSF). Poly-γ-glutamic acid (γ-PGA) was produced by Bacillus amyloliquefaciens C1 in SSF using dairy manure compost and monosodium glutamate production residuals as basic substrates. The production of γ-PGA reached a maximum of 0.6% after 20 days fermentation. Real-time polymerase chain reaction showed the amount of total bacteria reached 3.95 × 10(9) 16S rDNA copies/g sample after 30 days, which was in good accordance with the 4.80 × 10(9) CFU/g obtained by plate counting. Denaturing gradient gel electrophoresis profile showed a reduction of microbial diversity during fermentation, while the inoculum, B. amyloliquefaciens C1, was detected as the dominant organism through the whole process. In the mesophilic phase of SSF, Proteobacteria was the dominant microbial, which was replaced by Firmicutes and Actinobacteria in the thermophilic phase. The molecular analysis of the bacterial diversity has significant potential for instructing the maturing process of SSF to produce γ-PGA at a large-scale level, which could be a benefit in the production of high quality and stable SSF products.

摘要

细菌群落的动态在固态发酵(SSF)中起着重要作用。在 SSF 中,解淀粉芽孢杆菌 C1 以乳牛粪便堆肥和味精生产残渣为基本基质生产聚γ-谷氨酸(γ-PGA)。发酵 20 天后,γ-PGA 的产量达到最大值 0.6%。实时聚合酶链反应显示,30 天后总细菌数量达到 3.95×10(9)16S rDNA 拷贝/g 样品,与平板计数得到的 4.80×10(9)CFU/g 相符。变性梯度凝胶电泳图谱显示,发酵过程中微生物多样性减少,而接种物,解淀粉芽孢杆菌 C1,在整个过程中被检测为优势生物。在 SSF 的中温阶段,变形菌门是主要的微生物,在高温阶段被厚壁菌门和放线菌门取代。细菌多样性的分子分析对于指导 SSF 生产γ-PGA 的成熟过程具有重要意义,可以在大规模水平上生产高质量和稳定的 SSF 产品。

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