环境微生物中芳环加氧酶的获取策略

Discovery Strategies for Aromatic Oxygenases from Environmental Microbes

芳环加氧酶可催化芳香环的羟基化反应,广泛应用于生物修复及化工合成行业.本文在综述芳环加氧酶的分类和应用的基础上,详细探讨基于纯培养技术与宏基因组技术的芳环加氧酶开发策略,并首次对这些策略进行比较和评估.利用传统的纯培养技术已经获得大量的芳环加氧酶,然而这些加氧酶种类单一,重复率高,且新颖性不足.新兴的宏基因组技术赋予了开发未培养微生物资源的新思路,但利用其获取芳环加氧酶的研究刚刚起步,存在着筛选效率低、异源表达困难等难题.本文从基因信息量、筛选通量与效率、时间与成本以及目的基因下游表达可行性等方面对这两种方法进行综合评价,并提出宏基因组技术筛选不同类型芳环加氧酶的适用方法.纯培养技术与宏基因组技术的有机结合,与蓬勃发展的各种组学技术一起,将有助于推动芳环加氧酶的理论研究及生物催化产业的快速发展.

Aromatic oxygenases,which can catalyze the hydroxylation of aromatic ring,have been widely used in bioremediation and chemical synthesis.In this paper,the classification and application area of aromatic oxygenases are summarized.Moreover,their discovery strategies based on pure culture technology and metagenomic technology are discussed in detail and first evaluated.Abundant of aromatic oxygenases have been obtained using pure culture technology,but these enzymes exhibit with single type,high repetition rate and low novelty.Emerging metagenomic technology provides new ideas for discovering microbial resource from uncultured microbes.However,it is still in the initial stage of exploiting aromatic oxygenases by metagenomic methods,and many problems such as low screening efficiency and hard to express heterologously have not been resolved.This review evaluates the two strategies by analyzing the content of genetic information,screening throughout and efficiency,the time consume,cost,and expression feasibility of aromatic oxygenase.And the metagenomic methods suitable for screening different types of aromatic oxygenases are proposed.In the future,pure culture and metagenomic technology combined with other omics technologies,will contribute to promoting the theoretical study of aromatic oxygenases and rapid development of biocatalysis industry.