Engineering as a Cellulolytic Cell Factory for Production of -Coumaric Acid from Cellulose and Hemicellulose.

biosynthesis of high-value added food additive -coumaric acid (-CA) direct from cellulose/hemicellulose is a more sustainable route compared to the chemical route, considering the abundant cellulose/hemicellulose resources. In this study, a novel factory was constructed for the production of -CA in using cellulose/hemicellulose as the sole carbon source. Based on multicopy integration of the gene and reprogramming the shikimic acid pathway, the engineered strain produced 1035.

Characterization of a New Thermostable and Organic Solution-Tolerant Lipase from and Its Application in the Enrichment of Polyunsaturated Fatty Acids.

The search for and characterization of new lipases with excellent properties has always been urgent and is of great importance to meet industrial needs. In this study, a new lipase, from SBW25, belonging to the lipase subfamily I.3, was cloned and expressed in WB800N.

Copper Ion Mediates Yeast-to-Hypha Transition in .

Copper is an essential element that maintains yeast physiological function at low concentrations, but is toxic in excess. This study reported that Cu(II) significantly promoted the yeast-to-hypha transition of in dose-dependent manner. Strikingly, the intracellular Cu(II) accumulation was drastically reduced upon hyphae formation.

Employing Engineered Enolase Promoter for Efficient Expression of Lipase in via a Self-Excisable Vector.

is progressively being employed as a workhouse for recombinant protein expression. Here, we expanded the molecular toolbox by engineering the enolase promoter (pENO) and developed a new self-excisable vector, and based on this, a combined strategy was employed to enhance the expression of lipase (TLL) in . The strength of 11 truncated enolase promoters of different length was first identified using eGFP as a reporter.

A Novel Cre/-Based Genetic Tool for Repeated, Targeted and Markerless Gene Integration in .

The unconventional yeast is extensively applied in bioproduction fields owing to its excellent metabolite and protein production ability. Nonetheless, utilization of this promising host is still restricted by the limited availability of precise and effective gene integration tools. In this study, a novel and efficient genetic tool was developed for targeted, repeated, and markerless gene integration based on Cre/ site-specific recombination system.

High-Level Production of a Thermostable Mutant of Lipase 2 in .

As a promising biocatalyst, lipase 2 (YlLip2) is limited in its industrial applications due to its low thermostability. In this study, a thermostable YlLip2 mutant was overexpressed in and its half-life time was over 30 min at 80 °C. To obtain a higher protein secretion level, the gene dosage of the mutated gene was optimized and the lipase activity was improved by about 89%.

Overexpression of GRAS Rhizomucor miehei lipase in Yarrowia lipolytica via optimizing promoter, gene dosage and fermentation parameters.

Rhizomucor miehei lipase (RML), a GRAS catalyst with wide applications, was overexpressed in Yarrowia lipolytica, also a GRAS unconventional yeast, via a combined strategy, optimization for promoter, gene dosage and fermentation process. The lipase activity of the recombinant strain was first increased from 19.5 to 26.

Efficient gene disruption by posttransformational directed internal homologous recombination in Pichia pastoris.

In this study, we designed an improved method to construct gene-deficient strain in non-conventional yeast Pichia pastoris. This method achieved high efficiency of gene deletion without disruption of NHEJ-related genes. The disruption efficiency of Och1 reached about 80%.

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

This study is dedicated to efficiently produce lipase (ROL) by optimizing the expression of multiple expression-related helper proteins in . A series of engineered strains harboring different copy numbers of the gene and different copies of the chaperone gene were first constructed to examine the influence of gene copy number on ROL production. The results showed that multiple copies of gene did not significantly improve ROL expression.

New insight into the method of posttransformational vector amplification (PTVA) in Pichia pastoris.

Posttransformational vector amplification (PTVA) is widely used to enrich the gene-copy number in Pichia pastoris. We engineered two test strains for PTVA studies and demonstrate that the PTVA process results in the amplification of a fragment with the resistant gene flanked by two homologous arms instead of the entire vector.

High-level extracellular production of Rhizopus oryzae lipase in Pichia pastoris via a strategy combining optimization of gene-copy number with co-expression of ERAD-related proteins.

Rhizopus oryzae lipase (ROL) is an important industrial enzyme limited in application due to its low production in native strains. Here, we used a new combined strategy to overexpress ROL in Pichia pastoris. An efficient method based on bio-brick was developed to construct a series of vectors harboring different copy numbers of ROL gene cassettes, which were then transformed into P.

PmrA/PmrB Two-Component System Regulation of Expression in PAO1.

lipases are well-studied, but few studies have examined the mechanisms of lipase expression regulation. As a global regulatory protein, PmrA controls the expression of multiple genes such as the Dot/Icm apparatus, eukaryotic-like proteins, and secreted effectors. In this study, the effect of PmrA on expression of the lipase lipA in PAO1 was investigated by knocking out or overexpressing , and .

Engineering Yarrowia lipolytica to Simultaneously Produce Lipase and Single Cell Protein from Agro-industrial Wastes for Feed.

Lipases are scarcely exploited as feed enzymes in hydrolysis of lipids for increasing energy supply and improving nutrient use efficiency. In this work, we performed homologous overexpression, in vitro characterization and in vivo assessment of a lipase from the yeast Yarrowia lipolytica for feed purpose. Simultaneously, a large amount of yeast cell biomass was produced, for use as single cell protein, a potential protein-rich feed resource.

Preparation of Biodiesel with Liquid Synergetic Lipases from Rapeseed Oil Deodorizer Distillate.

To reduce industrial production cost, cheap and easily available rapeseed oil deodorizer distillates were used as feedstock to prepare biodiesel in this study. As a result, liquid forms of Candida rugosa lipase and Rhizopus oryzae lipase (ROL) were functioned as new and effective catalysts with biodiesel yield of 92.63% for 30 h and 94.

Overexpression of Candida rugosa lipase Lip1 via combined strategies in Pichia pastoris.

In this study, combined strategies were employed to heterologously overexpress Candida rugosa lipase Lip1 (CRL1) in a Pichia pastoris system. The LIP1 gene was systematically codon-optimized and synthesized in vitro. The Lip1 activity of a recombinant strain harboring three copies of the codon-optimized LIP1 gene reached 1200 U/mL in a shake flask culture.

Enhanced production of Thermomyces lanuginosus lipase in Pichia pastoris via genetic and fermentation strategies.

This study attempted to enhance the expression level of Thermomyces lanuginosus lipase (TLL) in Pichia pastoris using a series of strategies. The tll gene was first inserted into the expression vector pPIC9 K and transformed into P. pastoris strain GS115.