Engineering MtrAB two-component system enhances protein and glutamate export in corynebacterium glutamicum through cell wall remodeling.

Corynebacterium glutamicum serves as an attractive cell factory for protein secretion, surface display and glutamate production. Nevertheless, its complex cell wall structure imposes a bottleneck on biomolecule export. Although cell wall engineering has shown potential to overcome this limitation, how engineering cell wall biosynthesis affects the export of diverse biomolecules remains poorly understood.

Methanol metabolism in synthetic methylotrophic microorganisms.

Methanol has garnered increasing attention as an ideal carbon source for chemical biomanufacturing due to its abundance and easy availability. Microbial methylotrophy mainly refers to the ability to utilize reduced one-carbon compounds such as methanol, methane or formaldehyde as sole or primary sources of carbon and energy to sustain growth. Besides native methylotrophic microbes, such as Komagataella phaffii (K.

Modular deregulation of central carbon metabolism for efficient xylose utilization in Saccharomyces cerevisiae.

The tightly regulated central carbon metabolism in Saccharomyces cerevisiae, intricately linked to carbon sources utilized, poses a significant challenge to engineering efforts aimed at increasing the flux through its different pathways. Here, we present a modular deregulation strategy that enables high conversion rates of xylose through the central carbon metabolism. Specifically, employing a multifaceted approach encompassing five different engineering strategies-promoter engineering, transcription factor manipulation, biosensor construction, introduction of heterologous enzymes, and expression of mutant enzymes we engineer different modules of the central carbon metabolism at both the genetic and enzymatic levels.

Hybrid biological-chemical strategy for converting polyethylene into a recyclable plastic monomer using engineered Corynebacterium glutamicum.

Converting polyethylene (PE) into valuable materials, particularly ones that are better for the environment than the incumbent plastics, not only helps mitigate environmental issues caused by plastic waste but also alleviates the long-standing problem of microbial fermentation competing with food supplies. However, the inherent robustness of PE due to its strong carbon-carbon bonds and high molecular weight necessitates harsh decomposition conditions, resulting in diverse decomposition outcomes that present significant challenges for downstream applications, especially for bioconversion. In this study, we demonstrate a hybrid biological-chemical conversion process for PE, converting its decomposition products, namely short-chain diacids, into a monomer, β-keto-δ-lactone (BKDL), for highly recyclable polydiketoenimine plastics using engineered Corynebacterium glutamicum.

Enhanced methanol-xylose co-utilization strategy in Komagataella phaffii.

Bio-manufacturing based on non-food carbon sources is conducive to alleviating the global food crisis and greenhouse effect. However, the mechanism of the utilization of methanol and xylose in Komagataella phaffii based on endogenous metabolic pathways has not been fully explored. In this study, transcriptomics revealed a positive correlation between methanol metabolic efficiency and the transcription level of genes related to xylose metabolism and phosphate metabolism.

Maximizing Heterologous Expression of Engineered Type I Polyketide Synthases: Investigating Codon Optimization Strategies.

Type I polyketide synthases (T1PKSs) hold enormous potential as a rational production platform for the biosynthesis of specialty chemicals. However, despite great progress in this field, the heterologous expression of PKSs remains a major challenge. One of the first measures to improve heterologous gene expression can be codon optimization.

Creating NADP -Specific Formate Dehydrogenases from Komagataella phaffii by Enzymatic Engineering.

Most natural formate dehydrogenases (FDHs) exhibit NAD specificity, making it imperative to explore the engineering of FDH cofactor specificity for NADPH regeneration systems. The endogenous FDH of Komagataella phaffii (K. phaffii), termed KphFDH, is a typical NAD -specific FDH.

Improved polyketide production in C. glutamicum by preventing propionate-induced growth inhibition.

Corynebacterium glutamicum is a promising host for production of valuable polyketides. Propionate addition, a strategy known to increase polyketide production by increasing intracellular methylmalonyl-CoA availability, causes growth inhibition in C. glutamicum.

Strategies and challenges with the microbial conversion of methanol to high-value chemicals.

As alternatives to traditional fermentation substrates, methanol (CH OH), carbon dioxide (CO ) and methane (CH ) represent promising one-carbon (C1) sources that are readily available at low-cost and share similar metabolic pathway. Of these C1 compounds, methanol is used as a carbon and energy source by native methylotrophs, and can be obtained from CO and CH by chemical catalysis. Therefore, constructing and rewiring methanol utilization pathways may enable the use of one-carbon sources for microbial fermentations.

Glycerol transporter 1 (Gt1) and zinc-regulated transporter 1 (Zrt1) function in different modes for zinc homeostasis in Komagataella phaffii (Pichia pastoris).

Objectives: To identify the zinc transport function of the membrane proteins Gt1 and Zrt1 in Komagataella phaffii (Pichia pastoris) and study their regulatory mode.

Results: Two membrane proteins that might have zinc transport function were found in K. phaffii.

Characterization and application of a putative transcription factor (SUT2) in Pichia pastoris.

Pichia pastoris is able to metabolize methanol via a specific MUT (methanol utilization) pathway. Based on the powerful AOX1 (Alcohol Oxidase 1) promoter, the P. pastoris expression system has become one of the most widely used eukaryotic expression systems.

High efficiency CRISPR/Cas9 genome editing system with an eliminable episomal sgRNA plasmid in Pichia pastoris.

Pichia pastoris is a methylotrophic yeast in which host heterologous expression of proteins has been developed owing to the strong inducible alcohol oxidase promoter (P). However, it is difficult to manipulate the genome in P. pastoris.

Targeted editing of transcriptional activator MXR1 on the Pichia pastoris genome using CRISPR/Cas9 technology.

A highly efficient and targeted clustered regularly interspaced short palindromic repeats (CRISPR)/Cas9 gene editing system was constructed for Pichia pastoris (syn Komagataella phaffii). Plasmids containing single guide RNA and the methanol expression regulator 1 (MXR1) homology arms were used to precisely edit the transcriptional activator Mxr1 on the P. pastoris genome.

Transcriptional analysis of impacts of glycerol transporter 1 on methanol and glycerol metabolism in Pichia pastoris.

The efficient promoter of alcohol oxidase 1 (PAOX1) in methylotrophic yeast Pichia pastoris is strictly induced by methanol but repressed by glycerol with an unclear molecular mechanism. In the present study, the gene of a previously characterized transmembrane protein glycerol transporter 1 (GT1) of P. pastoris GS115 was deleted by homologous recombination.

Transcriptome analysis of Corynebacterium glutamicum in the process of recombinant protein expression in bioreactors.

Corynebacterium glutamicum (C. glutamicum) is a favorable host cell for the production of recombinant proteins, such as important enzymes and pharmaceutical proteins, due to its excellent potential advantages. Herein, we sought to systematically explore the influence of recombinant protein expression on the transcription and metabolism of C.

Transcription factor Mxr1 promotes the expression of Aox1 by repressing glycerol transporter 1 in Pichia pastoris.

In the methylotrophic yeast Pichia pastoris (P. pastoris), the efficient promoter of alcohol oxidase (PAox1) is induced by methanol and repressed by glycerol, but the molecular mechanism is not clear. In this study, the relationship between alcohol oxidase 1 (aox1), methanol expression regulator 1 (mxr1) and glycerol transporter 1 (gt1) was studied.

The Pichia pastoris transmembrane protein GT1 is a glycerol transporter and relieves the repression of glycerol on AOX1 expression.

Promoter of alcohol oxidase I (PAOX1) is the most efficient promoter involved in the regulation of recombinant protein expression in Pichia pastoris (P. pastoris). PAOX1 is tightly repressed by the presence of glycerol in the culture medium; thus, glycerol must be exhausted before methanol can be taken up by P.

Downsizing a pullulanase to a small molecule with improved soluble expression and secretion efficiency in Escherichia coli.

Background: Significant challenges, including low expression and extracellular secretion of soluble protein, are encountered in expressing and purifying Bacillus acidopullulyticus pullulanase (BaPul) in Escherichia coli.

Methods: An N-terminal domain truncation was adopted to facilitate BaPul variant expression and/or secretion.

Results: BaPul possesses a complex modular architecture that consists of CBM41-X45a-X25-X45b-CBM48-GH13.

WDR5 Expression Is Prognostic of Breast Cancer Outcome.

WDR5 is a core component of the human mixed lineage leukemia-2 complex, which plays central roles in ER positive tumour cells and is a major driver of androgen-dependent prostate cancer cell proliferation. Given the similarities between breast and prostate cancers, we explore the potential prognostic value of WDR5 gene expression on breast cancer survival. Our findings reveal that WDR5 over-expression is associated with poor breast cancer clinical outcome in three gene expression data sets and BreastMark.