Amnuaykanjanasin Alongkorn, Phonghanpot Suranat, Sengpanich Nattapong, Cheevadhanarak Supapon, Tanticharoen Morakot
National Center for Genetic Engineering and Biotechnology, National Science and Technology Development Agency, Pathumthani, Thailand.
Appl Environ Microbiol. 2009 Jun;75(11):3721-32. doi: 10.1128/AEM.02744-08. Epub 2009 Apr 3.
Polyketides draw much attention because of their potential use in pharmaceutical and biotechnological applications. This study identifies an abundant pool of polyketide synthase (PKS) genes from local isolates of tropical fungi found in Thailand in three different ecological niches: insect pathogens, marine inhabitants, and lichen mutualists. We detected 149 PKS genes from 48 fungi using PCR with PKS-specific degenerate primers. We identified and classified 283 additional PKS genes from 13 fungal genomes. Phylogenetic analysis of all these PKS sequences the comprising ketosynthase (KS) conserved region and the KS-acyltransferase interdomain region yielded results very similar to those for phylogenies of the KS domain and suggested a number of remarkable points. (i) Twelve PKS genes amplified from 12 different insect-pathogenic fungi form a tight cluster, although along with two PKS genes extracted from genomes of Aspergillus niger and Aspergillus terreus, in reducing clade III. Some of these insect-specific fungal PKSs are nearly identical. (ii) We identified 38 new PKS-nonribosomal peptide synthetase hybrid genes in reducing clade II. (iii) Four distinct clades were discovered with more than 75% bootstrap support. We propose to designate the novel clade D1 with 100% bootstrap support "reducing clade V." The newly cloned PKS genes from these tropical fungi should provide useful and diverse genetic resources for future research on the characterization of polyketide compounds synthesized by these enzymes.
聚酮化合物因其在制药和生物技术应用中的潜在用途而备受关注。本研究从泰国当地分离的热带真菌中鉴定出大量聚酮合酶(PKS)基因,这些真菌来自三个不同的生态位:昆虫病原体、海洋生物和地衣共生菌。我们使用PKS特异性简并引物通过PCR从48种真菌中检测到149个PKS基因。我们从13个真菌基因组中鉴定并分类了另外283个PKS基因。对所有这些包含酮合成酶(KS)保守区域和KS-酰基转移酶结构域间区域的PKS序列进行系统发育分析,结果与KS结构域的系统发育分析结果非常相似,并提出了一些值得注意的要点。(i)从12种不同的昆虫病原真菌中扩增出的12个PKS基因形成一个紧密的簇,尽管还有从黑曲霉和土曲霉基因组中提取的两个PKS基因,属于还原分支III。这些昆虫特异性真菌PKS中的一些几乎相同。(ii)我们在还原分支II中鉴定出38个新的PKS-非核糖体肽合成酶杂交基因。(iii)发现了四个不同的分支,自展支持率超过75%。我们建议将自展支持率为100%的新分支D1指定为“还原分支V”。从这些热带真菌中新克隆的PKS基因应为未来研究这些酶合成的聚酮化合物的特性提供有用且多样的遗传资源。