Optimization of Δ 9 -tetrahydrocannabinolic acid synthase production in Komagataella phaffii via post-translational bottleneck identification

Zirpel, Bastian; Degenhardt, Friederike; Zammarelli, Chantale; Wibberg, Daniel; Kalinowski, Jörn; Stehle, Felix; Kayser, Oliver

doi:10.1016/j.jbiotec.2018.03.008

Cited by 32 publications

(21 citation statements)

References 37 publications

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“…HAC1 can trigger the unfolded protein response (UPR) and increase the secretion of recombinant proteins which were demonstrated in many studies. For example, co-expression of HAC1 increased the expression of Δ9-Tetrahydrocannabinolic acid synthase 4.1-fold [17]. TRM1 is a transcription factor which is important for the transcriptional activation of genes of methanol utilization pathway in P. pastoris [18].…”

Section: Introductionmentioning

confidence: 99%

Multiple strategies to improve the yield of chitinase a from Bacillus licheniformis in Pichia pastoris to obtain plant growth enhancer and GlcNAc

et al. 2020

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Chitinase and chitin-oligosaccaride can be used in multiple field, so it is important to develop a high-yield chitinase producing strain. Here, a recombinant Pichia pastoris with 4 copies of ChiA gene from Bacillus licheniformis and co-expression of molecular chaperon HAC1 was constructed. The amount of recombinant ChiA in the supernatant of high-cell-density fermentation reaches a maximum of 12.7 mg/mL, which is 24-fold higher than that reported in the previous study. The recombinant ChiA can hydrolyze 30% collodidal chitin with 74% conversion ratio, and GlcNAc is the most abundant hydrolysis product, followed by N, N′-diacetylchitobiose. Combined with BsNagZ, the hydrolysate of ChiA can be further transformed into GlcNAc with 88% conversion ratio. Additionally, the hydrolysate of ChiA can obviously accelerate the germination growth of rice and wheat, increasing the seedling height and root length by at least 1.6 folds within 10 days.

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Section: Introductionmentioning

confidence: 99%

Multiple strategies to improve the yield of chitinase a from Bacillus licheniformis in Pichia pastoris to obtain plant growth enhancer and GlcNAc

et al. 2020

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“…For example, the secretion of glucose oxidase (GOD) was remarkably enhanced via co-expression of Sec53 [ 20 ]. However, the overexpression of Sec53 did not increase the production of Δ 9 -tetrahydrocannabinolic acid synthase [ 27 ]. Thus, the effectiveness of these strategies for the expression of a specific protein, including ROL, may be uncertain.…”

Section: Introductionmentioning

confidence: 99%

Efficient Heterologous Production of Rhizopus oryzae Lipase via Optimization of Multiple Expression-Related Helper Proteins

Jiao

Zhou

et al. 2018

IJMS

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This study is dedicated to efficiently produce Rhizopus oryzae lipase (ROL) by optimizing the expression of multiple expression-related helper proteins in Pichia pastoris. A series of engineered strains harboring different copy numbers of the ROL gene and different copies of the chaperone Pdi gene were first constructed to examine the influence of Pdi gene copy number on ROL production. The results showed that multiple copies of Pdi gene did not significantly improve ROL expression. Then, the effect of the co-overexpression of 10 expression-related helper proteins on ROL secretion was investigated by screening 20 colonies of each transformants. The data from shaking-flask fermentation suggested that Ssa4, Bmh2, Sso2, Pdi, Bip, Hac1, and VHb had positive effects on ROL expression. Subsequently, Ssa4, Bmh2, and Sso2, which all participate in vesicular trafficking and strongly promote ROL expression, were combined to further improve ROL production level. ROL activity of the screened strain GS115/5ROL-Ssa4-Sso2-Bmh2 4# attained 5230 U/mL. Furthermore, when the helper proteins Pdi, Bip, Hac1, and VHb were individually co-expressed with ROL in the strain GS115/5ROL-Ssa4-Sso2-Bmh2 4#, lipase activity increased to 5650 U/mL in the strain GS115/5ROL-Ssa4-Sso2-Bmh2-VHb 9#. Additionally, the maximum ROL activity of 41,700 U/mL was achieved in a 3 L bioreactor for high-density fermentation via a sorbitol–methanol co-feeding strategy, reaching almost twofold the value of the initial strain GS115/pAOα-5ROL 11#. Thus, the strategies in this study significantly increased ROL expression level, which is of great potential for the large-scale production of ROL in P. pastoris.

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“…It is hypothesized that CsaCHIL, a chalcone isomerase-like protein lacking catalytic activity, potentially binds THCA and/or CBDA for stabilization in hemp glandular trichomes and limits negative feedback to upstream enzymes. It has also been shown that upregulation of multiple foldases and chaperones resulted in a 20-fold improvement of THCA synthase functionality in yeast and poses a promising avenue for optimizing microbial production [20].…”

Section: Recent Advances In Microbial Transgene Expression Systems Fomentioning

confidence: 99%

New frontiers: harnessing pivotal advances in microbial engineering for the biosynthesis of plant-derived terpenoids

Belcher

Mahinthakumar

Keasling

2020

Current Opinion in Biotechnology

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Optimization of Δ 9 -tetrahydrocannabinolic acid synthase production in Komagataella phaffii via post-translational bottleneck identification

Cited by 32 publications

References 37 publications

Multiple strategies to improve the yield of chitinase a from Bacillus licheniformis in Pichia pastoris to obtain plant growth enhancer and GlcNAc

Multiple strategies to improve the yield of chitinase a from Bacillus licheniformis in Pichia pastoris to obtain plant growth enhancer and GlcNAc

Efficient Heterologous Production of Rhizopus oryzae Lipase via Optimization of Multiple Expression-Related Helper Proteins

New frontiers: harnessing pivotal advances in microbial engineering for the biosynthesis of plant-derived terpenoids

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