角蛋白降解能力差异菌株筛选体系的建立

Establishment of screening system for strains with different keratin degradation ability

以微生物降解为主的羽毛生物炼制是资源化利用其的重要手段之一,但机理不明是限制其进一步应用的瓶颈。突变体库筛选性状变异菌株的正向遗传学是研究分子机理的常用方案,该方案要求高效可靠的筛选方法。然而,目前依靠测酶活方法来决定降解能力,费时费力,难以用于大规模筛选。本文通过优化羽毛培养基成分及接种菌浓度,建立了一种用于筛选不同角蛋白降解能力微生物的筛选体系。利用该体系分析由mini Tn10构建的S. maltophinia DHHJ突变体库,得到了6株羽毛角蛋白降解能力减弱的突变体,筛选结果与酶活测试100%一致。该研究结果为寻找与角蛋白降解相关基因奠定了基础。

Feather biorefinery is one of the important means of resource utilization, but the unclear degrading mechanism is the bottleneck of its further application. The forward genetics of mutant library screening mutant strains is a common scheme to study the molecular mechanism, which requires efficient and reliable screening methods. However, at present, it is time-consuming and laborious to determine the degradation ability by measuring enzyme activity, and it is difficult to be used for large-scale screening. In this paper, a screening system for screening microorganisms with different keratin degradation ability was established by optimizing the composition of the feather medium and the concentration of the inoculum. The S. maltophinia DHHJ mutant library constructed by mini Tn 10 was analyzed by this system, and six mutants with weakened degradation ability of feather keratin were obtained. The screening results were consistent with the enzyme activity test. The results of this study laid a foundation for the search for genes related to keratin degradation.