Constitutive hyperproduction of sorbicillinoids in Trichoderma reesei ZC121.
Constitutive hyperproduction of sorbicillinoids in Trichoderma reesei ZC121.
复制标题
里氏木霉 ZC121 中山梨西林类化合物的组成型过量生产
DOI：
10.1186/s13068-018-1296-4
复制
发表时间：
2018
影响因子：
6.3
通讯作者：
Chen Z
中科院分区：
文献类型：
--
作者：
Li C;Lin F;Sun W;Yuan S;Zhou Z;Wu FG;Chen Z
背景除了其突出的纤维素酶生产能力外，里氏木霉还产生多种有价值的次生代谢产物，但其生产迄今为止尚未受到太多关注。其中，山梨西林类化合物是一大类源自聚酮化合物的六酮化合物次级代谢产物，由于其表现出多种重要的生物功能，包括抗癌、抗氧化、抗病毒和抗菌特性，正引起研究人员越来越多的兴趣。因此，开发具有山梨西林类化合物的组成型、超量生产的真菌菌株是未来工业应用所期望的，但尚未得到充分研究。此外，虽然里氏木霉已被证明能够产生山梨西林类化合物，并具有相应的基因簇和提出的生物合成途径，但控制山梨西林类化合物生物合成的潜在分子机制仍不清楚。结果考虑到脱靶，通过在里氏木霉第四号染色体端粒上插入基因12121敲除盒，从菌株RUT-C30构建重组里氏木霉ZC121在基因 121121 不成功删除过程中遇到诱变。当在纤维素上生长时，菌株 ZC121 显示纤维素酶产量急剧减少，但与菌株 RUT-C30 相比，山梨西林类化合物产量显着增强。山梨西林类化合物的过量生产是一个组成过程，与碳源、光、pH 和温度等培养条件无关。据我们所知，菌株 ZC121 在葡萄糖和纤维素上生长时显示出创纪录的山梨西林生产水平。山梨西醇和双维替诺龙是生产的两种主要山梨西林类化合物。与 RUT-C30 相比，ZC121 显示出不同的形态并显着减少孢子形成，但具有相似的生长速率和生物量。转录组分析表明，在纤维素上生长的 ZC121 中，大多数参与纤维素酶生产的基因显着下调，而山梨西林类基因簇中的所有基因在纤维素和葡萄糖上均上调。山梨西林类化合物的生产记录。里氏木霉首次降解纤维素，以高产率生产蛋白质以外的平台化合物。我们认为，如 ZC121 的表型分析和比较转录组分析所示，端粒区域发生的脱靶诱变可能会导致染色体重塑，从而改变菌株 ZC121 的细胞结构和整体基因表达模式。总体而言，里氏木霉ZC121对于山梨西林类化合物的工业化生产具有广阔的前景，并且可以作为探索山梨西林类化合物生物合成调控机制的良好模型。
BackgroundIn addition to its outstanding cellulase production ability, Trichoderma reesei produces a wide variety of valuable secondary metabolites, the production of which has not received much attention to date. Among them, sorbicillinoids, a large group of hexaketide secondary metabolites derived from polyketides, are drawing a growing interest from researchers because they exhibit a variety of important biological functions, including anticancer, antioxidant, antiviral, and antimicrobial properties. The development of fungi strains with constitutive, hyperproduction of sorbicillinoids is thus desired for future industry application but is not well-studied. Moreover, although T. reesei has been demonstrated to produce sorbicillinoids with the corresponding gene cluster and biosynthesis pathway proposed, the underlying molecular mechanism governing sorbicillinoid biosynthesis remains unknown.ResultsRecombinant T. reesei ZC121 was constructed from strain RUT-C30 by the insertion of the gene 12121-knockout cassette at the telomere of T. reesei chromosome IV in consideration of the off-target mutagenesis encountered during the unsuccessful deletion of gene 121121. Strain ZC121, when grown on cellulose, showed a sharp reduction of cellulase production, but yet a remarkable enhancement of sorbicillinoids production as compared to strain RUT-C30. The hyperproduction of sorbicillinoids is a constitutive process, independent of culture conditions such as carbon source, light, pH, and temperature. To the best of our knowledge, strain ZC121 displays record sorbicillinoid production levels when grown on both glucose and cellulose. Sorbicillinol and bisvertinolone are the two major sorbicillinoid compounds produced. ZC121 displayed a different morphology and markedly reduced sporulation compared to RUT-C30 but had a similar growth rate and biomass. Transcriptome analysis showed that most genes involved in cellulase production were downregulated significantly in ZC121 grown on cellulose, whereas remarkably all genes in the sorbicillinoid gene cluster were upregulated on both cellulose and glucose.ConclusionA constitutive sorbicillinoid-hyperproduction strain T. reesei ZC121 was obtained by off-target mutagenesis, displaying an overwhelming shift from cellulase production to sorbicillinoid production on cellulose, leading to a record for sorbicillinoid production. For the first time, T. reesei degraded cellulose to produce platform chemical compounds other than protein in high yield. We propose that the off-target mutagenesis occurring at the telomere region might cause chromosome remodeling and subsequently alter the cell structure and the global gene expression pattern of strain ZC121, as shown by phenotype profiling and comparative transcriptome analysis of ZC121. Overall, T. reesei ZC121 holds great promise for the industrial production of sorbicillinoids and serves as a good model to explore the regulation mechanism of sorbicillinoids’ biosynthesis.
登录
查看更多内容
DOI：
10.1007/s10482-017-0952-1
发表时间：
2018-03
期刊：
Antonie van Leeuwenhoek
影响因子：
--
作者：
Aerts D;Hauer EE;Ohm RA;Arentshorst M;Teertstra WR;Phippen C;Ram AFJ;Frisvad JC;Wösten HAB
通讯作者：
Wösten HAB
影响因子：
6.3
作者：
Häkkinen M;Valkonen MJ;Westerholm-Parvinen A;Aro N;Arvas M;Vitikainen M;Penttilä M;Saloheimo M;Pakula TM
通讯作者：
Pakula TM
影响因子：
32.5
作者：
Brethauer, Simone;Studer, Michael Hanspeter
通讯作者：
Studer, Michael Hanspeter
影响因子：
4.9
作者：
Abe, N;Hirota, A
通讯作者：
Hirota, A
影响因子：
5.2
作者：
Derntl C;Guzmán-Chávez F;Mello-de-Sousa TM;Busse HJ;Driessen AJM;Mach RL;Mach-Aigner AR
通讯作者：
Mach-Aigner AR