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Minjie Gao,Zhongping Shi,Shijuan Dong,Ruisong Yu,Jianrong Wu,Zhiyong Zheng,Xiaobei Zhan 한국화학공학회 2011 Korean Journal of Chemical Engineering Vol.28 No.6
The performance of traditional heterologous protein production by Pichia pastoris with methanol induction at 30 ℃ is poor, characterized by low ATP regeneration rate and weak operation stability. A low temperature induction strategy at 20 ℃ was thus adopted for efficient porcine interferon-α production in a 10 L fermentor. With the strategy,maximal methanol tolerance level could reach about 40 g/L to effectively deal with methanol concentration variations,so that the complicated on-line methanol measurement system could be eliminated. Moreover, metabolic analysis based on multiple state-variables measurements indicated that pIFN-α antiviral activity enhancement profited from the formation of an efficient ATP regeneration system at 20℃ induction. Compared to the induction strategy at 30 ℃, the proposed strategy increased the ATP regeneration rate by 49-66%, the maximal p_IFN-α antiviral activity was enhanced about 20-fold and reached a higher level of 1.5×10^6 IU/mL.
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Xin Liu,Yichen Jiang,Yinghui Zhang,Mingzheng Yu,Hongjun Jiang,Jianhong Xu,Jianrong Shi 한국미생물학회 2019 The journal of microbiology Vol.57 No.8
Dihydroxyacid dehydratase (DHAD), encoded by ILV3, catalyses the third step in the biosynthetic pathway of branchedchain amino acids (BCAAs), which include isoleucine (Ile), leucine (Leu), and valine (Val). Enzymes involved in BCAA biosynthesis exist in bacteria, plants, and fungi but not in mammals and are therefore attractive targets for antimicrobial or herbicide development. In this study, three paralogous ILV3 genes (FgILV3A, FgILV3B, and FgILV3C) were identified in the genome of Fusarium graminearum, the causal agent of Fusarium head blight (FHB). Deletion of FgILV3A alone or combined with FgILV3B or FgILV3C indicated an important role for FgILV3A in BCAA biosynthesis. FgILV3A deletion mutants lost the ability to grow on medium lacking amino acids. Exogenous supplementation of 1 mM Ile and Val rescued the auxotrophy of ΔFgIlv3A, though 5 mM was required to recover the growth defects in ΔFgIlv3AB and ΔFgIlv3AC strains, indicating that FgIlv3b and FgIlv3c exhibit redundant but accessory roles with FgIlv3a in BCAA biosynthesis. The auxotrophy of ΔFgIlv3A resulted in pleiotropic defects in aerial hyphal growth, in conidial formation and germination, and in aurofusarin accumulation. In addition, the mutants showed reduced virulence and deoxynivalenol production. Overall, our study demonstrates that FgIlv3a is crucial for BCAA biosynthesis in F. graminearum and a candidate fungicide target for FHB management.
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