He Ronglin, Li Chen, Ma Lijuan, Zhang Dongyuan, Chen Shulin
Tianjin Institute of Industrial Biotechnology, Chinese Academy of Sciences, Tianjin, 300308, People's Republic of China.
Key Laboratory of Industrial Fermentation Microbiology, Ministry of Education, Tianjin University of Science and Technology, Tianjin, 300457, People's Republic of China.
3 Biotech. 2016 Dec;6(2):214. doi: 10.1007/s13205-016-0516-5. Epub 2016 Oct 5.
The morphology of Trichoderma reesei is a vitally important factor for cellulase productivity. This study investigated the effect of hyphal morphology on cellulase production in the hyper-cellulolytic mutant, T. reesei DES-15. With a distinct morphology, T. reesei DES-15 was obtained through Diethyl sulfite (DES) mutagenesis. The hyphal morphology of DES-15 batch-cultured in a 5-L fermentor was significantly shorter and more branched than the parental strain RUT C30. The cellulase production of DES-15 during batch fermentation was 66 % greater than that of RUT C30 when cultured the same conditions. DES-15 secreted nearly 50 % more protein than RUT C30. The gene expression level of a set of genes (cla4, spa2, ras2, ras1, rhoA, cdc42, and racA) known to be involved in hyphae growth and hyphal branching was measured by quantitative real-time PCR. The transcriptional analysis of these genes demonstrated that a decrease in gene expressions might contribute to the increased hyphal branching seen in DES-15. These results indicated that the highly branching hyphae in DES-15 resulted in increased cellulase production, suggesting that DES-15 may be a good candidate for use in the large-scale production of cellulase.
里氏木霉的形态是纤维素酶产量的一个至关重要的因素。本研究调查了菌丝形态对纤维素酶高产突变体里氏木霉DES-15纤维素酶产生的影响。通过亚硫酸二乙酯(DES)诱变获得了具有独特形态的里氏木霉DES-15。在5-L发酵罐中分批培养的DES-15的菌丝形态比亲本菌株RUT C30明显更短且分支更多。在相同条件下培养时,DES-15在分批发酵过程中的纤维素酶产量比RUT C30高66%。DES-15分泌的蛋白质比RUT C30多近50%。通过定量实时PCR测量了一组已知参与菌丝生长和菌丝分支的基因(cla4、spa2、ras2、ras1、rhoA、cdc42和racA)的基因表达水平。这些基因的转录分析表明,基因表达的降低可能导致了DES-15中观察到的菌丝分支增加。这些结果表明,DES-15中高度分支的菌丝导致纤维素酶产量增加,这表明DES-15可能是大规模生产纤维素酶的良好候选菌株。