A5103046645- Microbial Metabolic Engineering and Bioproduction
- Bacterial Genetics and Biotechnology
- CRISPR and Genetic Engineering
- RNA and protein synthesis mechanisms
- Enzyme Catalysis and Immobilization
- Biofuel production and bioconversion
- Bacteriophages and microbial interactions
- biodegradable polymer synthesis and properties
- Photosynthetic Processes and Mechanisms
- Microplastics and Plastic Pollution
- Porphyrin Metabolism and Disorders
- Genomics and Phylogenetic Studies
- Plant biochemistry and biosynthesis
- Gene Regulatory Network Analysis
- Enzyme Structure and Function
- Viral Infectious Diseases and Gene Expression in Insects
- Fungal and yeast genetics research
- Probiotics and Fermented Foods
- Enzyme Production and Characterization
- Biochemical Acid Research Studies
- Glycosylation and Glycoproteins Research
- Amino Acid Enzymes and Metabolism
- Escherichia coli research studies
- Biochemical and Molecular Research
- DNA Repair Mechanisms
Shandong University
2016-2025
State Key Laboratory of Microbial Technology
2016-2025
Qingdao Binhai University
2022-2024
Chinese Academy of Sciences
2018-2022
Qingdao Institute of Bioenergy and Bioprocess Technology
2018-2022
Qingdao Center of Resource Chemistry and New Materials
2020
Tsinghua University
2015
Leipzig University
2007
Poly(ethylene terephthalate) (PET) is the most abundant polyester plastic and causing serious environmental pollution. Rapid biological depolymerization of PET waste at a large scale requires powerful engineered enzymes with excellent performance. Here, we designed computational strategy to analyze ligand affinity energy chains by molecular docking dynamic protein conformations, named analysis based on (ADD). After three rounds engineering assisted ADD, drastically enhanced...
Abstract Phenolic compounds are industrially versatile chemicals, also the most ubiquitous pollutants. Recently, biosynthesis and biodegradation of phenols has attracted increasing attention, while phenols’ toxicity is a major issue. Here, we evolved phloroglucinol-tolerant Escherichia coli strains via adaptive evolution, three mutations (Δ sodB , Δ clpX fetAB overexpression) prove great assistance in tolerance improvement. We discover that phloroglucinol complexes with iron promotes...
The QuikChange™ site-directed mutagenesis method is popular but imperfect. An improvement by using partially overlapping primers has been reported several times; however, it incompatible with the proposed mechanism. complementary to linearly amplify a target plasmid products annealing produce double-stranded DNA molecules 5'-overhangs. overhang supposed form circular plasmids staggered breaks, which can be repaired in Escherichia coli after transformation. Here, we demonstrated that PCR...
Abstract Homologous recombination-mediated genome engineering has been broadly applied in prokaryotes with high efficiency and accuracy. However, this method is limited realizing larger-scale editing numerous genes or large DNA fragments because of the relatively complicated procedure for template construction. Here, we describe a CRISPR-Cas9 assisted non-homologous end-joining (CA-NHEJ) strategy rapid efficient inactivation bacterial gene (s) homologous recombination-independent manner...
Abstract Background Yarrowia lipolytica , a non-traditional oil yeast, has been widely used as platform for lipid production. However, the production of other chemicals such terpenoids in engineered Y. is still low. α-Farnesene, sesquiterpene, can be medicine, bioenergy and fields, very high economic value. Here, we α-farnesene an example to explore potential terpenoid Results We constructed libraries strains overexpressing mevalonate pathway synthase genes by non-homologous end-joining...
The management and recycling of plastic waste is a challenging global issue. Polyethylene terephthalate (PET), one the most widely used synthetic plastics, can be hydrolyzed by series enzymes. However, upcycling resulting monomers also problem. In this study, we designed co-cultivation system, in which PET degradation was coupled with polyhydroxybutyrate (PHB) production. First, PETase from Ideonalla sakaiensis expressed Yarrowia lipolytica Po1f signal peptide lipase. engineered...
The oleaginous yeast Yarrowia lipolytica represents an environmentally friendly platform cell factory for β-carotene production. However, Y. is a dimorphic species that can undergo yeast-to-mycelium transition when exposed to stress. mycelial form unfavorable industrial fermentation. In this study, β-carotene-producing strains were constructed via the integration of multiple copies 13 genes related biosynthesis pathway. content increased by 11.7-fold compared with start strain T1. As...
Polyethylene terephthalate (PET) has caused serious environmental concerns but could be degraded at high temperature. Previous studies show that cutinase from Thermobifida fusca KW3 (TfCut2) is capable of degrading and upcycling PET limited by its thermal stability. Nowadays, Popular protein stability modification methods rely mostly on the crystal structures, ignore fact actual conformation complex constantly changing. To solve these problems, we developed a computational approach to design...
Abstract Succinic acid (SA) is an important C4-dicarboxylic acid. Microbial production of SA at low pH results in purification costs and hence good overall process economics. However, redox imbalances limited biosynthesis from glucose via the reductive tricarboxylic (TCA) cycle yeast. Here, we engineer strictly aerobic yeast Yarrowia lipolytica for efficient without control. Introduction TCA into cytosol a succinate dehydrogenase-disrupted strain causes arrested cell growth. Although...
In order to obtain functional expression of PHA synthase gene phaC2 from Pseudomonas aeruginosa in Escherichia coli, the coding region was subcloned, including ribosomal binding site, into pBluescript SK- collinear lac promoter. This plasmid pBHR71-C2 enabled E. coli LS1298 (fadB) under promoter control, leading accumulation, when grown LB medium containing 0.5% (w/v) various fatty acids (C8-C14). The strongest accumulation observed, dodecanoate provided as carbon source, and contributed 15%...
Lactobacillus plantarum is a food-grade microorganism with industrial and medical relevance belonging to the group of lactic acid bacteria (LAB). Traditional strategies for obtaining gene deletion variants in this organism are mainly vector-based double-crossover methods, which inefficient laborious. A feasible possibility solve problem recombineering, greatly expands possibilities engineering DNA molecules vivo various organisms. In work, double-stranded (dsDNA) recombineering system was...
Abstract Direct optimization of the metabolic pathways on chromosome requires tools that can fine tune overexpression a desired gene or optimize combination multiple genes. Although plasmid-dependent has been used for this task, fundamental issues concerning its genetic stability and operational repeatability have not addressed. Here, we describe rapid reliable strategy chromosomal integration gene(s) with copies (CIGMC), which uses flippase from yeast 2-μm plasmid. Using green fluorescence...
Corynebacterium glutamicum is generally regarded as a safe microorganism and used to produce many biochemicals, including L-glutamate. 5-Aminolevulinic acid (ALA) an L-glutamate derived non-protein amino acid, widely applied in fields such medicine agriculture.The products of the gltX, hemA, hemL genes participate synthesis ALA from Their annotated C. homologs were shown be functional using heterologous complementation overexpression techniques. Coexpression hemA native host led accumulation...
CRISPR/Cas9-assisted recombineering is restricted in lactic acid bacteria because of the lack available antibiotics and vectors. In this study, a seamless genome editing method was carried out Lactobacillus plantarum using double-stranded DNA (dsDNA) single-stranded (ssDNA) recombineering, recombination efficiency effectively improved by endogenous adenine-specific methyltransferase overexpression. L. WCFS1 produced 797.3 mg/liter N -acetylglucosamine (GlcNAc) through reinforcement GlcNAc...
β-Farnesene is a sesquiterpene with versatile industrial applications. The production of β-farnesene from waste lipid feedstock an attractive method for sustainable and recycling oil. Yarrowia lipolytica unconventional oleaginous yeast, which can use has great potential to synthesize acetyl-CoA-derived chemicals.
With the rapid development of synthetic biology, oleaginous yeast Yarrowia lipolytica has become an attractive microorganism for chemical production. To better optimize and reroute metabolic pathways, we have expanded CRISPR-based gene expression toolkit Y. lipolytica. By sorting integration sites associated with high expression, new neutral efficiency were identified. Diverse genetic components, including promoters terminators, also characterized to expand range. We found that in addition...
Abstract Yarrowia lipolytica is an important oleaginous industrial microorganism used to produce biofuels and other value‐added compounds. Although several genetic engineering tools have been developed for Y. , there no efficient method genomic integration of large DNA fragments. In addition, methods constructing multigene expression libraries biosynthetic pathway optimization are still lacking in . this study, we demonstrate that multiple fragments can be randomly efficiently integrated...