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| Enrichment Culturing and Bacterial Community Structures Analysis of a Cold-adapted Lignocellulose Degrading Microflora |
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Abstract In the face of the inefficiency of bioconversion of agricultural biomass waste in low temperature environment in the winter, this study was aimed at enrichment culture and community structure analysis of a cold-adapted cellulose degrading microflora A25-3, which was isolated from the soil in alpine region. The microflora A25-3 could grow rapidly by enrichment culture at 15 ℃, with 10% cow dung and 10% LB sodium salt, and the weight loss ratio of cow dung was 48.28% in 72 h. The carboxymethyl cellulose (CMCase) activity of the supernatant of microflora culture at 15 ℃, with a peak of 42.37 U in 120 h was higher than that at 25 ℃. Total microbial DNA was directly extracted from the microflora cultured at different temperatures of 15 and 25 ℃. The clone library of 16S rRNA genes was amplified using PCR with universal bacterial primer sets. Each unique restriction fragment polymorphism pattern, which was created with 2 restriction endonucleases (HinfⅠand Csp6Ⅰ), was designated as an operational taxonomic unit (OTU). Amplified DNA was used for diversity analysis. The microflora phylogenetic trees of bacterial clone library at different temperatures of 15 and 25 ℃ was constructed. Community structure analysis revealed that the genera Brevundimonas, Pseudomonas, Chryseobacterium, Acinetobacte, Sphingobacterium and Bacillus were dominant in the clone library from microflora cultured at 25 ℃, while the genera Acinetobacte, Psychrobacillus and Bacillus were dominant at 15 ℃. The results suggested that the dominant population of cold-adapted cellulose degrading bacteria important species, especially Acinetobacte, provided the effect of increasing the CMCase activity at 15 ℃. The result will provide a scientific basis for the next study to separate active bacteria strains from the bacterial community and to optimize cellulase synthesis conditions.
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Received: 30 December 2014
Published: 13 April 2015
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