Review
Biotechnology & Applied Microbiology
Hang-Qin Zhu, Xiao-Ling Tang, Ren-Chao Zheng, Yu-Guo Zheng
Summary: By specifically incorporating UAAs into proteins through genetic code expansion, more structural and functional features can be endowed to proteins, which have been widely utilized in enzyme engineering. This method provides opportunities for tuning and expanding the functional properties of enzymes.
WORLD JOURNAL OF MICROBIOLOGY & BIOTECHNOLOGY
(2021)
Review
Biotechnology & Applied Microbiology
Pritam Giri, Amol D. Pagar, Mahesh D. Patil, Hyungdon Yun
Summary: This review discusses various strategies of chemical modification of enzymes, including traditional methods utilizing reactivities of canonical amino acids and emerging techniques like noncanonical amino acid-mediated 'click' chemistry and dehydroalanine (Dha)-mediated modifications, to improve functional properties and stability of industrially important biocatalysts.
BIOTECHNOLOGY ADVANCES
(2021)
Review
Biochemistry & Molecular Biology
Ella Watkins-Dulaney, Sabine Straathof, Frances Arnold
Summary: Tryptophan synthase is a paragon of noncanonical amino acid synthesis and is ideal for synthetic and biological applications. Directed evolution has enabled significant expansion of the enzyme's substrate scope, allowing for improved efficiency in biocatalytic cascades.
Article
Chemistry, Applied
Andrea Varga, Pal Csuka, Orlavanah Sonesouphap, Gergely Banoczi, Monica Ioana Tosa, Gabriel Katona, Zsofia Molnar, Laszlo Csaba Bencze, Laszlo Poppe, Csaba Paizs
Summary: A novel phenylalanine ammonia-lyase (PzaPAL) was identified from the psychrophilic yeast Pseudozyma antarctica, showing significantly different substrate binding pocket compared to known PALs. Recombinant PzaPAL exhibited higher turnover numbers than PcPAL and demonstrated enhanced enantiotope selectivity in both kinetic resolutions and ammonia addition reactions.
Article
Biophysics
Elnaz Zeynaloo, Elsayed M. Zahran, Yu-Ping Yang, Emre Dikici, Trajen Head, Leonidas G. Bachas, Sylvia Daunert
Summary: This study introduces a new method of using unnatural amino acids as protein labels, which not only changes the local microenvironment but also has minimal impact on the overall structure. By incorporating L-DOPA into the protein, it can be used as an electrochemical label for highly selective and sensitive detection of glucose.
BIOSENSORS & BIOELECTRONICS
(2022)
Article
Chemistry, Physical
Yaoyun Wu, Sheng Zhang, Wei Song, Jia Liu, Xiulai Chen, Guipeng Hu, Yiwen Zhou, Liming Liu, Jing Wu
Summary: The catalytic efficiency of enzymatic synthesis of alpha-keto acids from alpha-amino acids deamination is limited by the catalytic distance between the substrate and the cofactor. Through protein engineering strategy, two variants were developed to shorten the distance and improve the efficiency of the process. The modified variants showed improved substrate binding pose and led to successful conversion of selected substrates with high yields.
Article
Chemistry, Multidisciplinary
James L. Galman, Fabio Parmeggiani, Lisa Seibt, William R. Birmingham, Nicholas J. Turner
Summary: In this study, an enzyme cascade system was constructed by engineering enzymes and cell strains, which could produce high-value chemicals from lignin.
ANGEWANDTE CHEMIE-INTERNATIONAL EDITION
(2022)
Article
Biotechnology & Applied Microbiology
Xinjie Chen, Yingying Liu, Jiaqi Hou, Yuan Lu
Summary: The study proposed a cell-free unnatural protein synthesis system based on linear DNA templates for rapid high-throughput screening and evolution, selecting optimal OTS components for UNAAs. Screening results showed that the o-aaRS and o-tRNA of pPaF demonstrated good orthogonality, while a new pair for pBpF was identified, demonstrating the efficiency of this method in accelerating the screening process for versatile UNAAs.
SYNTHETIC AND SYSTEMS BIOTECHNOLOGY
(2021)
Article
Chemistry, Multidisciplinary
Yan Li, Junqing Wang, Fen Wang, Li Wang, Leilei Wang, Ziqi Xu, Haibo Yuan, Xiaohui Yang, Piwu Li, Jing Su, Ruiming Wang
Summary: The study modified the beta-oxidation pathway of Escherichia coli and designed a P450 terminal hydroxylase (CYP153A33-CPRBM3) for the synthesis of 10-HDA, achieving a yield of 217 mg L-1 from decanoic acid.
Article
Chemistry, Applied
Yosuke Sumida, Masayuki Yamasaki, Yoshiaki Nishiya, Shinya Kumagai, Toshihide Yamada, Masayuki Azuma
Summary: This study screened DSA mutants for improved enantioselectivity and thermal stability through directed evolution, with the L182E mutant showing superior performance. The crystal structure of the L182E mutant was solved, providing insight into its improved thermostability. The coupled enzymatic method involving L182E DSA and NSAR demonstrated high conversion rates and enantiomeric excesses for d-phenylalanine and d-tryptophan production, suggesting the potential industrial application of L182E DSA as a biocatalyst for d-amino acids production.
ADVANCED SYNTHESIS & CATALYSIS
(2021)
Review
Chemistry, Multidisciplinary
Pooja Laxman, Shirin Ansari, Katharina Gaus, Jesse Goyette
Summary: Single Molecule Localization Microscopy (SMLM) is an imaging method that allows visualization of structures smaller than the diffraction limit of light. Incorporating unnatural amino acids (UAAs) with attached fluorescent dyes for protein labeling enables precise localization and visualization of individual molecules. Comparing this method with other protein-based labeling techniques, the site-specific incorporation of UAAs coupled with fluorescent dyes in SMLM has both benefits and shortcomings.
FRONTIERS IN CHEMISTRY
(2021)
Review
Biotechnology & Applied Microbiology
Roland Wohlgemuth, Jennifer Littlechild
Summary: The biosynthesis of metabolites is increasingly important for various applications in the life sciences. Enzymes in biological systems can synthesize these molecules under sustainable conditions, and modular cell-free biocatalytic systems offer opportunities for their synthesis.
FRONTIERS IN BIOENGINEERING AND BIOTECHNOLOGY
(2022)
Article
Chemistry, Multidisciplinary
Ludwig L. Bengel, Benjamin Aberle, Alexander-N. Egler-Kemmerer, Samuel Kienzle, Bernhard Hauer, Stephan C. Hammer
Summary: Selective alkylation of pyrazoles using a catalyst-controlled enzymatic system allows for the efficient synthesis of important molecules with unprecedented regioselectivity. The use of a computational enzyme-library design tool enabled the conversion of a promiscuous enzyme into a small enzyme family of pyrazole-alkylating enzymes in one round of mutagenesis and screening. This enzymatic system demonstrates high selectivity and achieves different alkylations of pyrazoles with remarkable regiodivergence.
ANGEWANDTE CHEMIE-INTERNATIONAL EDITION
(2021)
Article
Nanoscience & Nanotechnology
Zeynep Demirsoy, Gulcihan Gulseren
Summary: Self-assembling enzyme mimics are an effective system for imitating the activity of natural enzymes. By tuning intermolecular and intramolecular interactions, efficient enzyme mimics can be formed. These designed nanocatalysts can mimic different enzyme classes and exhibit catalytic activity under physiological conditions.
ACS APPLIED NANO MATERIALS
(2022)
Article
Biochemical Research Methods
Alexander Virklund, Christian Bille Jendresen, Alex Toftgaard Nielsen, John M. Woodley
Summary: The characteristics of TAL enzyme extracted from Flavobacterium johnsoniae and Herpetosiphon aurantiacus were studied. It was found that FjTAL has higher activity but weaker inhibition, and enzyme engineering is required to reduce inhibition and improve stability.
BIOTECHNOLOGY JOURNAL
(2023)
Article
Biotechnology & Applied Microbiology
Geon-Hee Kim, Hyunwoo Jeon, Taresh P. Khobragade, Mahesh D. Patil, Sihyong Sung, Sanghan Yoon, Yumi Won, In Suk Choi, Hyungdon Yun
ENZYME AND MICROBIAL TECHNOLOGY
(2019)
Article
Chemistry, Physical
Sung-Yeon Joo, Hee-Wang Yoo, Sharad Sarak, Byung-Gee Kim, Hyungdon Yun
Article
Chemistry, Physical
Sanghan Yoon, Mahesh D. Patil, Sharad Sarak, Hyunwoo Jeon, Geon-Hee Kim, Taresh P. Khobragade, Sihyong Sung, Hyungdon Yun
Article
Multidisciplinary Sciences
Sunghark Kwon, Jun Hyuck Lee, Chang Min Kim, Hyunseok Jang, Hyungdon Yun, Ju-Hong Jeon, Insuk So, Hyun Ho Park
SCIENTIFIC REPORTS
(2019)
Article
Chemistry, Physical
Mahesh D. Patil, Sanghan Yoon, Hyunwoo Jeon, Taresh P. Khobragade, Sharad Sarak, Amol D. Pagar, Yumi Won, Hyungdon Yun
Review
Biotechnology & Applied Microbiology
Yumi Won, Amol D. Pagar, Mahesh D. Patil, Philip E. Dawson, Hyungdon Yun
BIOTECHNOLOGY AND BIOPROCESS ENGINEERING
(2019)
Article
Chemistry, Physical
Sharad Sarak, Hyunwoo Jeon, Mahesh D. Patil, Taresh P. Khobragade, Amol D. Pagar, Sihyong Sung, Hee-Wang Yoo, Byung-Gee Kim, Sung Ho Yoon, Hyungdon Yun
Article
Agriculture, Multidisciplinary
Sanghwan Ko, Hyunwoo Jeon, Sanghan Yoon, Munsu Kyung, Hyungdon Yun, Jung-Hyun Na, Sang Taek Jung
JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY
(2020)
Article
Chemistry, Multidisciplinary
Sharad Sarak, Sihyong Sung, Hyunwoo Jeon, Mahesh D. Patil, Taresh P. Khobragade, Amol D. Pagar, Philip E. Dawson, Hyungdon Yun
Summary: A highly atom-efficient integrated cofactor/co-product recycling cascade using cycloalkylamines for the synthesis of nylon building blocks was reported, achieving excellent conversions with E. coli whole cells and purified enzymes. The tandem biocatalytic cascade was demonstrated to produce lactams using engineered biocatalysts, showing promise as an alternative for bio-based production of omega-amino acids and corresponding lactam compounds.
ANGEWANDTE CHEMIE-INTERNATIONAL EDITION
(2021)
Review
Chemistry, Multidisciplinary
Amol D. Pagar, Mahesh D. Patil, Dillon T. Flood, Tae Hyeon Yoo, Philip E. Dawson, Hyungdon Yun
Summary: The two main strategies for enzyme engineering, directed evolution and rational design, have been widely used to improve the intrinsic activities of proteins. However, the limited chemical diversity of biopolymers, restricted to 20 canonical amino acids, hinders the creation of novel enzymes. To overcome this, researchers have been devoted to expanding the protein sequence space through chemical modifications and/or incorporation of noncanonical amino acids.
Article
Chemistry, Multidisciplinary
Hee-Wang Yoo, Hyunsang Jung, Sharad Sarak, Ye Chan Kim, Beom Gi Park, Byung-Gee Kim, Mahesh D. Patil, Hyungdon Yun
Summary: In this study, multi-enzymatic cascade reaction systems using Escherichia coli-based cell modules were designed to generate biopolymer monomers. These systems provide a greener alternative for the biosynthesis of various biopolymer monomers from renewable saturated fatty acids.
Article
Chemistry, Multidisciplinary
Uk-Jae Lee, Junghyeon Ko, Su-Hwan Kim, Pyung-Gang Lee, Young-Hyeon An, Hyungdon Yun, Dillon T. Flood, Philip E. Dawson, Nathaniel S. Hwang, Byung-Gee Kim
Summary: This study presents a method for in situ biosynthesis of artificial melanin in skin using photoactivatable tyrosinase. By introducing a photocleavable o-nitrobenzyl tyrosine into the tyrosinase, its enzymatic activity can be recovered through photo-cleavage. Using this method, melanin particles and hydrogel can be synthesized in the skin for tissue protection. Encapsulating the photoactivatable tyrosinase into lipid nanoparticles allows transdermal delivery into the skin tissue.
Article
Chemistry, Multidisciplinary
Sharad Sarak, Taresh P. Khobragade, Hyunwoo Jeon, Amol D. Pagar, Pritam Giri, Somin Lee, Hyungdon Yun
Summary: A multienzyme biocatalytic cascade was developed to synthesize nylon monomers with high efficiency and a self-sufficient redox network, utilizing a cell-based modularization strategy to produce 6-aminohexanoic acid and epsilon-caprolactam from cyclohexanol. The designed E. coli consortia showed high yields of the desired products, making it a cost-effective method for nylon precursor production.
Article
Chemistry, Multidisciplinary
Yumi Won, Hyunwoo Jeon, Amol D. Pagar, Mahesh D. Patil, Saravanan Prabhu Nadarajan, Dillon T. Flood, Philip E. Dawson, Hyungdon Yun
CHEMICAL COMMUNICATIONS
(2019)