Elimination of the Sugar Transporter GAT1 Increased Xylanase I Production in Trichoderma reesei

Wei, Xu; Yu, Fang; Ma, Ding; Ren, Yuxin; Meng, Xiangfeng; Chen, Guanjun; Zhang, Weixin; Liu, Weifeng

doi:10.3389/fmicb.2022.810066

Cited by 2 publications

(2 citation statements)

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“…Meanwhile, sugar transporters are indispensable for substrate exchange between the cytoplasm and environment and can regulate cellulase induction, probably by transporting inducers or working as signal transceptors [14]. Although 55 sugar transporters have been annotated in the T. reesei genome [31], only a few of them have been characterized [16,19,[32][33][34]. Among these sugar transporters, the cellobiose/lactose transporter CRT1 is critical for cellulase induction in T. reesei, and the disruption of CRT1 impaired cellulase induction either on cellulose or soluble lactose [14][15][16].…”

Section: Discussionmentioning

confidence: 99%

Role of cellulose response transporter-like protein CRT2 in cellulase induction in Trichoderma reesei

Su¹,

Xu²,

Xiang³

2023

Biotechnol Biofuels

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Background Induction of cellulase in cellulolytic fungi Trichoderma reesei is strongly activated by cellulosic carbon sources. The transport of cellulosic inducer and the perception of inducing signal is generally considered as the critical process for cellulase induction, that the inducing signal would be perceived by a sugar transporter/transceptor in T. reesei. Several sugar transporters are coexpressed during the induction stage, but which function they serve and how they work collaboratively are still difficult to elucidate. Results In this study, we found that the constitutive expression of the cellulose response transporter-like protein CRT2 (previously identified as putative lactose permease TRE77517) improves cellulase induction on a cellulose, cellobiose or lactose medium. Functional studies indicate that the membrane-bound CRT2 is not a transporter of cellobiose, lactose or glucose in a yeast system, and it also does not affect cellobiose and lactose utilization in T. reesei. Further study reveals that CRT2 has a slightly similar function to the cellobiose transporter CRT1 in cellulase induction. Overexpression of CRT2 led to upregulation of CRT1 and the key transcription factor XYR1. Moreover, overexpression of CRT2 could partially compensate for the function loss of CRT1 on cellulase induction. Conclusions Our study uncovers the novel function of CRT2 in cellulase induction collaborated with CRT1 and XYR1, possibly as a signal transductor. These results deepen the understanding of the influence of sugar transporters in cellulase production.

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Section: Discussionmentioning

confidence: 99%

Role of cellulose response transporter-like protein CRT2 in cellulase induction in Trichoderma reesei

Su¹,

Xu²,

Xiang³

2023

Biotechnol Biofuels

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“…SDS-PAGE and western blot were performed according to standard protocols. XYNI and XYNII were immunoblotted using polyclonal antibodies raised against peptides of XYNI (amino acids of 52 ~ 65) and XYNII (amino acids of 79 ~ 92) [ 34 ], respectively.…”

Section: Methodsmentioning

confidence: 99%

A novel transcriptional repressor specifically regulates xylanase gene 1 in Trichoderma reesei

Xu,

Ren,

Xia

et al. 2023

Biotechnol Biofuels

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Background The well-known industrial fungus Trichoderma reesei has an excellent capability of secreting a large amount of cellulases and xylanases. The induced expression of cellulase and xylanase genes is tightly controlled at the transcriptional level. However, compared to the intensive studies on the intricate regulatory mechanism of cellulase genes, efforts to understand how xylanase genes are regulated are relatively limited, which impedes the further improvement of xylanase production by T. reesei via rational strain engineering. Results To identify transcription factors involved in regulating xylanase gene expression in T. reesei, yeast one-hybrid screen was performed based on the promoters of two major extracellular xylanase genes xyn1 and xyn2. A putative transcription factor named XTR1 showing significant binding capability to the xyn1 promoter but not that of xyn2, was successfully isolated. Deletion of xtr1 significantly increased the transcriptional level of xyn1, but only exerted a minor promoting effect on that of xyn2. The xylanase activity was increased by ~ 50% with XTR1 elimination but the cellulase activity was hardly affected. Subcellular localization analysis of XTR1 fused to a green fluorescence protein demonstrated that XTR1 is a nuclear protein. Further analyses revealed the precise binding site of XTR1 and nucleotides critical for the binding within the xyn1 promoter. Moreover, competitive EMSAs indicated that XTR1 competes with the essential transactivator XYR1 for binding to the xyn1 promoter. Conclusions XTR1 represents a new transcriptional repressor specific for controlling xylanase gene expression. Isolation and functional characterization of this new factor not only contribute to further understanding the stringent regulatory network of xylanase genes, but also provide important clues for boosting xylanase biosynthesis in T. reesei.

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Elimination of the Sugar Transporter GAT1 Increased Xylanase I Production in Trichoderma reesei

Cited by 2 publications

References 50 publications

Role of cellulose response transporter-like protein CRT2 in cellulase induction in Trichoderma reesei

Role of cellulose response transporter-like protein CRT2 in cellulase induction in Trichoderma reesei

A novel transcriptional repressor specifically regulates xylanase gene 1 in Trichoderma reesei

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