Network


Latest external collaboration on country level. Dive into details by clicking on the dots.

Hotspot


Dive into the research topics where Ee Lui Ang is active.

Publication


Featured researches published by Ee Lui Ang.


Metabolic Engineering | 2016

A highly efficient single-step, markerless strategy for multi-copy chromosomal integration of large biochemical pathways in Saccharomyces cerevisiae

Shuobo Shi; Youyun Liang; Mingzi M. Zhang; Ee Lui Ang; Huimin Zhao

Despite recent advances in genome editing capabilities for the model organism Saccharomyces cerevisiae, the chromosomal integration of large biochemical pathways for stable industrial production remains challenging. In this work, we developed a simple platform for high-efficiency, single-step, markerless, multi-copy chromosomal integration of full biochemical pathways in Saccharomyces cerevisiae. In this Di-CRISPR (delta integration CRISPR-Cas) platform based on the Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR) and CRISPR-associated systems (Cas), we specifically designed guide RNA sequences to target multiple delta sites in the yeast genome. The generation of double stranded breaks at the delta sites allowed simultaneous integration of multiple copies of linearized donor DNA containing large biochemical pathways. With our newly developed Di-CRISPR platform, we were able to attain highly efficient and markerless integration of large biochemical pathways and achieve an unprecedented 18-copy genomic integration of a 24 kb combined xylose utilization and (R,R)-2,3-butanediol (BDO) production pathway in a single step, thus generating a strain that was able to produce BDO directly from xylose. The simplicity and high efficiency of the Di-CRISPR platform could provide a superior alternative to high copy plasmids and would render this platform an invaluable tool for genome editing and metabolic engineering in S. cerevisiae.


Nature Chemical Biology | 2017

CRISPR-Cas9 strategy for activation of silent Streptomyces biosynthetic gene clusters

Mingzi M. Zhang; Fong Tian Wong; Yajie Wang; Shangwen Luo; Yee Hwee Lim; Elena Heng; Wan Lin Yeo; Ryan E. Cobb; Behnam Enghiad; Ee Lui Ang; Huimin Zhao

Here we report an efficient CRISPR-Cas9 knock-in strategy to activate silent biosynthetic gene clusters (BGCs) in streptomycetes. We applied this one-step strategy to activate multiple BGCs of different classes in five Streptomyces species and triggered the production of unique metabolites, including a novel pentangular type II polyketide in Streptomyces viridochromogenes. This potentially scalable strategy complements existing activation approaches and facilitates discovery efforts to uncover new compounds with interesting bioactivities.


Drug Design Development and Therapy | 2015

Recent advances in combinatorial biosynthesis for drug discovery.

Huihua Sun; Zihe Liu; Huimin Zhao; Ee Lui Ang

Because of extraordinary structural diversity and broad biological activities, natural products have played a significant role in drug discovery. These therapeutically important secondary metabolites are assembled and modified by dedicated biosynthetic pathways in their host living organisms. Traditionally, chemists have attempted to synthesize natural product analogs that are important sources of new drugs. However, the extraordinary structural complexity of natural products sometimes makes it challenging for traditional chemical synthesis, which usually involves multiple steps, harsh conditions, toxic organic solvents, and byproduct wastes. In contrast, combinatorial biosynthesis exploits substrate promiscuity and employs engineered enzymes and pathways to produce novel “unnatural” natural products, substantially expanding the structural diversity of natural products with potential pharmaceutical value. Thus, combinatorial biosynthesis provides an environmentally friendly way to produce natural product analogs. Efficient expression of the combinatorial biosynthetic pathway in genetically tractable heterologous hosts can increase the titer of the compound, eventually resulting in less expensive drugs. In this review, we will discuss three major strategies for combinatorial biosynthesis: 1) precursor-directed biosynthesis; 2) enzyme-level modification, which includes swapping of the entire domains, modules and subunits, site-specific mutagenesis, and directed evolution; 3) pathway-level recombination. Recent examples of combinatorial biosynthesis employing these strategies will also be highlighted in this review.


Applied and Environmental Microbiology | 2014

Engineered Pentafunctional Minicellulosome for Simultaneous Saccharification and Ethanol Fermentation in Saccharomyces cerevisiae

Youyun Liang; Tong Si; Ee Lui Ang; Huimin Zhao

ABSTRACT Several yeast strains have been engineered to express different cellulases to achieve simultaneous saccharification and fermentation of lignocellulosic materials. However, successes in these endeavors were modest, as demonstrated by the relatively low ethanol titers and the limited ability of the engineered yeast strains to grow using cellulosic materials as the sole carbon source. Recently, substantial enhancements to the breakdown of cellulosic substrates have been observed when lytic polysaccharide monooxygenases (LPMOs) were added to traditional cellulase cocktails. LPMOs are reported to cleave cellulose oxidatively in the presence of enzymatic electron donors such as cellobiose dehydrogenases. In this study, we coexpressed LPMOs and cellobiose dehydrogenases with cellobiohydrolases, endoglucanases, and β-glucosidases in Saccharomyces cerevisiae. These enzymes were secreted and docked onto surface-displayed miniscaffoldins through cohesin-dockerin interaction to generate pentafunctional minicellulosomes. The enzymes on the miniscaffoldins acted synergistically to boost the degradation of phosphoric acid swollen cellulose and increased the ethanol titers from our previously achieved levels of 1.8 to 2.7 g/liter. In addition, the newly developed recombinant yeast strain was also able to grow using phosphoric acid swollen cellulose as the sole carbon source. The results demonstrate the promise of the pentafunctional minicellulosomes for consolidated bioprocessing by yeast.


Scientific Reports | 2016

Metabolic engineering of a synergistic pathway for n-butanol production in Saccharomyces cerevisiae

Shuobo Shi; Tong Si; Zihe Liu; Hongfang Zhang; Ee Lui Ang; Huimin Zhao

n-Butanol has several favourable properties as an advanced fuel or a platform chemical. Bio-based production of n-butanol is becoming increasingly important for sustainable chemical industry. Synthesis of n-butanol can be achieved via more than one metabolic pathway. Here we report the metabolic engineering of Saccharomyces cerevisiae to produce n-butanol through a synergistic pathway: the endogenous threonine pathway and the introduced citramalate pathway. Firstly, we characterized and optimized the endogenous threonine pathway; then, a citramalate synthase (CimA) mediated pathway was introduced to construct the synergistic pathway; next, the synergistic pathway was optimized by additional overexpression of relevant genes identified previously; meanwhile, the n-butanol production was also improved by overexpression of keto-acid decarboxylases (KDC) and alcohol dehydrogenase (ADH). After combining these strategies with co-expression of LEU1 (two copies), LEU4, LEU2 (two copies), LEU5, CimA, NFS1, ADH7 and ARO10*, we achieved an n-butanol production of 835 mg/L in the final engineered strain, which is almost 7-fold increase compared to the initial strain. Furthermore, the production showed a 3-fold of the highest titer ever reported in yeast. Therefore, the engineered yeast strain represents a promising alternative platform for n-butanol production.


Expert Opinion on Drug Discovery | 2017

Using natural products for drug discovery: the impact of the genomics era

Mingzi M. Zhang; Yuan Qiao; Ee Lui Ang; Huimin Zhao

ABSTRACT Introduction: Evolutionarily selected over billions of years for their interactions with biomolecules, natural products have been and continue to be a major source of pharmaceuticals. In the 1990s, pharmaceutical companies scaled down their natural product discovery programs in favor of synthetic chemical libraries due to major challenges such as high rediscovery rates, challenging isolation, and low production titers. Propelled by advances in DNA sequencing and synthetic biology technologies, insights into microbial secondary metabolism provided have inspired a number of strategies to address these challenges. Areas covered: This review highlights the importance of genomics and metagenomics in natural product discovery, and provides an overview of the technical and conceptual advances that offer unprecedented access to molecules encoded by biosynthetic gene clusters. Expert opinion: Genomics and metagenomics revealed nature’s remarkable biosynthetic potential and her vast chemical inventory that we can now prioritize and systematically mine for novel chemical scaffolds with desirable bioactivities. Coupled with synthetic biology and genome engineering technologies, significant progress has been made in identifying and predicting the chemical output of biosynthetic gene clusters, as well as in optimizing cluster expression in native and heterologous host systems for the production of pharmaceutically relevant metabolites and their derivatives.


ACS Synthetic Biology | 2017

A New Era of Genome Integration—Simply Cut and Paste!

Zihe Liu; Youyun Liang; Ee Lui Ang; Huimin Zhao

Genome integration is a powerful tool in both basic and applied biological research. However, traditional genome integration, which is typically mediated by homologous recombination, has been constrained by low efficiencies and limited host range. In recent years, the emergence of homing endonucleases and programmable nucleases has greatly enhanced integration efficiencies and allowed alternative integration mechanisms such as nonhomologous end joining and microhomology-mediated end joining, enabling integration in hosts deficient in homologous recombination. In this review, we will highlight recent advances and breakthroughs in genome integration methods made possible by programmable nucleases, and their new applications in synthetic biology and metabolic engineering.


Bioorganic & Medicinal Chemistry | 2017

Biocatalysis for the synthesis of pharmaceuticals and pharmaceutical intermediates

Huihua Sun; Hongfang Zhang; Ee Lui Ang; Huimin Zhao

Biocatalysis has been increasingly used for pharmaceutical synthesis in an effort to make manufacturing processes greener and more sustainable. Biocatalysts that possess excellent activity, specificity, thermostability and solvent-tolerance are highly sought after to meet the requirements of practical applications. Generating biocatalysts with these specific properties can be achieved by either discovery of novel biocatalysts or protein engineering. Meanwhile, chemoenzymatic routes have also been designed and developed for pharmaceutical synthesis on an industrial scale. This review discusses the recent discoveries, engineering, and applications of biocatalysts for the synthesis of pharmaceuticals and pharmaceutical intermediates. Key classes of biocatalysts include reductases, oxidases, hydrolases, lyases, isomerases, and transaminases.


Chemcatchem | 2016

Production of Adipic Acid from Sugar Beet Residue by Combined Biological and Chemical Catalysis

Hongfang Zhang; Xiukai Li; Xiaoyun Su; Ee Lui Ang; Yugen Zhang; Huimin Zhao

Adipic acid is one of the most important industrial dicarboxylic acids and is used mainly as a precursor to nylon‐6,6. Currently, commercial adipic acid is produced primarily from benzene by a chemical route that is associated with environmental, health, and safety concerns. Herein, we report a new process to produce adipic acid from an inexpensive renewable feedstock, sugar beet residue by combining an engineered Escherichia coli strain and Re‐based chemical catalysts. The engineered E. coli converted d‐galacturonic acid to mucic acid, which was precipitated easily with acid, and the mucic acid was further converted to adipic acid by a deoxydehydration reaction catalyzed by an oxorhenium complex followed by a Pt/C‐catalyzed hydrogenation reaction under mild conditions. A high selectivity to the free acid products was achieved by tuning the acidity of the Re‐based catalysts. Finally, adipic acid was produced directly from sugar beet residue that was hydrolyzed enzymatically with engineered E. coli and two chemical catalysts in a yield of 8.4 %, which signifies a new route for the production of adipic acid.


Current Opinion in Chemical Biology | 2018

Recent developments in the application of P450 based biocatalysts

Yifeng Wei; Ee Lui Ang; Huimin Zhao

Cytochrome P450 monooxygenases (P450s) catalyze regioselective and stereoselective oxidative modifications of a wide variety of substrates, and are involved in the biosynthesis of many natural products. Despite the complex requirements of the P450 reaction system and its intransigence to recombinant expression, the promise of harnessing P450 reactivity for the industrial-scale production of specialty chemicals has led to much effort invested in P450 engineering. Here we review recent developments (between 2015 and 2017) in the application of P450s and their engineered variants as biocatalysts. We describe strategies for the reconstitution of P450 activity in heterologous microbial hosts, and the expanding repertoire of non-natural reactions catalyzed by engineered P450s.

Collaboration


Dive into the Ee Lui Ang's collaboration.

Top Co-Authors

Avatar
Top Co-Authors

Avatar
Top Co-Authors

Avatar
Top Co-Authors

Avatar
Top Co-Authors

Avatar
Top Co-Authors

Avatar
Top Co-Authors

Avatar
Top Co-Authors

Avatar
Top Co-Authors

Avatar
Top Co-Authors

Avatar
Researchain Logo
Decentralizing Knowledge