为获得产卡拉胶酶并高效降解麒麟菜的菌株,降解麒麟菜以利于其中有效成分的释放,以卡拉胶为唯一碳源,通过透明圈法进行初筛,DNS法酶活力测定和发酵麒麟菜降解率测定进行复筛,从南海的活藻石中筛选到1株卡拉胶酶活力和麒麟菜降解率较高的菌株ZN-8,经形态学观察、生理生化试验和16S rDNA序列分析,确定该菌为海洋嗜纤维菌属(Cellulophaga sp.)。在单因素试验基础上,通过Plackett-Burman试验确定影响菌株发酵降解麒麟菜的主要因素[发酵温度、(NH4)2SO4添加量、发酵时间]。通过响应面优化试验建立回归方程,得到该菌株发酵麒麟菜最佳条件:接种量10%,(NH4)2SO4添加量0.5 g/L,料液比20(g∶L),发酵温度27 ℃,发酵时间6.5 d,发酵初始pH值为8。在最佳发酵条件下,麒麟菜降解率达到48.93%,相比于优化前提高了36.79%。综上,菌株ZN-8在麒麟菜降解方面具有较大潜力,通过对其发酵麒麟菜条件进行优化,显著提高了其降解效率,对于促进麒麟菜的绿色加工和高值化利用具有重要意义,为麒麟菜资源的开发提供了思路和参考。
In the quest for a strain of producing carrageenanase and efficiently degrading Eucheuma to facilitate the release of its active ingredients, this study used carrageenan as the sole carbon source to prepare culture medium.This study isolated one strain ZN-8 with high carrageenanase activity and degradation rate of Eucheuma from the living algae stones in the South China Sea by screening with transparent circle method, re-screening by enzyme activity assay of DNS method and degradation rate of fermented Eucheuma.The strain was identified as Cellulophaga sp.through morphological observation, physiological and biochemical tests, and 16S rDNA sequence 下0.analysis.Based on the single-factor test, the main factors affecting the degradation of Eucheuma were determined by the Plackett-Burman test [fermentation temperature, (NH4)2SO4 addition, fermentation time].The regression equation was established by response surface test, and the optimal conditions included 10% inoculum, 0.5 g/L (NH4)2SO4, 20 g/L material-liquid ratio, 27 ℃, 6.5 days, and the initial pH 8.Under the optimal fermentation conditions, the degradation rate of Eucheuma reached 48.93%, which was a 36.79% improvement over pre-optimization levels.In conclusion, the strain ZN-8 showed significant potential for Eucheuma degradation, enhancing green processing and high-value utilization, providing insights for resource development of Eucheuma.
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