Macdonald Spencer S, Pereira Jose H, Liu Feng, Tegl Gregor, DeGiovanni Andy, Wardman Jacob F, Deutsch Samuel, Yoshikuni Yasuo, Adams Paul D, Withers Stephen G
Michael Smith Laboratories, University of British Columbia, 2185 East Mall, Vancouver, British Columbia V6T 1Z4, Canada.
Department of Chemistry, University of British Columbia, 2036 Main Mall, Vancouver, British Columbia V6T 1Z1, Canada.
ACS Cent Sci. 2022 Apr 27;8(4):430-440. doi: 10.1021/acscentsci.1c01570. Epub 2022 Mar 16.
The considerable utility of glycoside phosphorylases (GPs) has led to substantial efforts over the past two decades to expand the breadth of known GP activities. Driven largely by the increase of available genomic DNA sequence data, the gap between the number of sequences in the carbohydrate active enzyme database (CAZy DB) and its functionally characterized members continues to grow. This wealth of sequence data presented an exciting opportunity to explore the ever-expanding CAZy DB to discover new GPs with never-before-described functionalities. Utilizing an sequence analysis of CAZy family GH94, we discovered and then functionally and structurally characterized the new GP β-1,3--acetylglucosaminide phosphorylase. This new GP was sourced from the genome of the cell-wall-less Mollicute bacterium, and was found to synthesize β-1,3-linked -acetylglucosaminide linkages. The resulting poly-β-1,3--acetylglucosamine represents a new, previously undescribed biopolymer that completes the set of possible β-linked GlcNAc homopolysaccharides together with chitin (β-1,4) and PNAG (poly-β-1,6--acetylglucosamine). The new biopolymer was denoted , a combination of the genus and the polysaccharide chitin, and the new GP was thus denoted acholetin phosphorylase (AchP). Use of the reverse phosphorolysis action of AchP provides an efficient method to enzymatically synthesize acholetin, which is a new biodegradable polymeric material.
在过去二十年中，糖苷磷酸化酶（GPs）的显著实用性促使人们做出了大量努力来扩展已知GP活性的范围。在很大程度上受到可用基因组DNA序列数据增加的推动，碳水化合物活性酶数据库（CAZy DB）中的序列数量与其功能表征成员之间的差距持续扩大。这些丰富的序列数据为探索不断扩展的CAZy DB提供了一个令人兴奋的机会，以发现具有前所未有的功能的新型GPs。通过对CAZy家族GH94进行序列分析，我们发现并对新型GPβ-1,3-乙酰氨基葡萄糖磷酸化酶进行了功能和结构表征。这种新型GP源自无细胞壁的柔膜菌属细菌的基因组，被发现可合成β-1,3连接的乙酰氨基葡萄糖键。由此产生的聚-β-1,3-乙酰氨基葡萄糖代表了一种新的、以前未描述过的生物聚合物，它与几丁质（β-1,4）和PNAG（聚-β-1,6-乙酰氨基葡萄糖）一起构成了所有可能的β连接的GlcNAc同多糖。这种新的生物聚合物被命名为，它是柔膜菌属和多糖几丁质的组合，因此这种新型GP被命名为阿乔letin磷酸化酶（AchP）。利用AchP的反向磷酸解作用提供了一种酶促合成阿乔letin的有效方法，阿乔letin是一种新型可生物降解的聚合材料。
