糖质的体外生物合成技术及应用进展

朱玥明; 杨建刚; 曾艳; 董乾震; 孙媛霞

doi:10.12211/2096-8280.2025-050

您当前的位置：

首页 >

文章列表页 >

糖质的体外生物合成技术及应用进展

特约评述 | 更新时间：2026-09-04

- 糖质的体外生物合成技术及应用进展
- Advances and applications of in vitro biosynthesis of carbohydrates
- 合成生物学 2026年7卷第4期页码：781-797
- 作者机构：
  
  1.中国科学院天津工业生物技术研究所，天津 300308
  2.合成生物学海河实验室，天津 300308
  3.国家合成生物技术创新中心，天津 300308
  4.低碳合成工程生物学全国重点实验室，天津 300308
- 作者简介：
  
  朱玥明（1982—），男，博士，副研究员。研究方向为酶工程、微生物代谢工程。E-mail：zhu_ym@tib.cas.cn
  孙媛霞（1963—），女，博士，研究员。研究方向为糖生物学、糖工程与酶工程，建立了功能糖类及其衍生物的绿色生物制造技术。E-mail：sun_yx@tib.cas.cn
- 基金信息：
  
  中国科学院关键核心技术攻坚先导专项(XDC0120203);天津合成生物技术创新能力提升行动(TSBICIP-KJGG-028);山东省重点研发计划项目(2023CXGC010714);合成生物学海河实验室颠覆性创新项目(22HHSWSS00016);合成生物学海河实验室攻关项目(22HHSWSS00003)
- DOI：10.12211/2096-8280.2025-050
  中图分类号： Q81
- 收稿：2025-05-28，
  
  修回：2025-07-08，
  
  纸质出版：2026-08-31
- 稿件说明：
移动端阅览
朱玥明，杨建刚，曾艳，董乾震，孙媛霞. 糖质的体外生物合成技术及应用进展［J］. 合成生物学， 2026， 7（4）： 781-797

ZHU Yueming， YANG Jian′gang， ZENG Yan， DONG Qianzhen， SUN Yuanxia. Advances and applications of in vitro biosynthesis of carbohydrates［J］. Synthetic Biology Journal， 2026， 7（4）： 781-797
朱玥明，杨建刚，曾艳，董乾震，孙媛霞. 糖质的体外生物合成技术及应用进展［J］. 合成生物学， 2026， 7（4）： 781-797 DOI： 10.12211/2096-8280.2025-050.

ZHU Yueming， YANG Jian′gang， ZENG Yan， DONG Qianzhen， SUN Yuanxia. Advances and applications of in vitro biosynthesis of carbohydrates［J］. Synthetic Biology Journal， 2026， 7（4）： 781-797 DOI： 10.12211/2096-8280.2025-050.

摘要

糖质是功能食品、医药、能源与材料领域中至关重要的生物资源。近年来，作为一种新兴的无细胞合成平台，体外生物合成技术正逐渐成为糖质制造的重要技术路径。该技术通过整合多种酶在单一反应体系中协同催化，突破了细胞底盘的稳态限制，具备更高的路径可控性、反应效率与原子经济性。本文系统回顾了糖质体外合成体系的核心技术进展，包括新酶挖掘与分子改造、热力学驱动与辅酶循环策略、模块化路径设计及多酶协同适配机制；同时梳理了该技术在单糖/糖醇、寡糖、多糖及糖衍生物等方面的代表性应用案例，展现其在廉价资源利用、原子经济性及规模化生产方面的优势。此外，本文还分析了人工智能在新酶挖掘改造、反应路径优化、工艺参数预测等糖质体外生物合成关键技术中的应用潜力。随着生物学与众多学科的深度交叉融合，糖质体外生物合成正加速从实验室走向工业化，有望成为下一代绿色生物制造的重要支撑。

Abstract

Carbohydrates are indispensable bioresources with diverse and profound applications across the realms of functional foods

pharmaceuticals

energy

and advanced materials. The traditional methods for carbohydrate production

which typically rely on cellular systems

often suffer from inherent limitations such as low product yield

complex pathway regulation

and limited scalability. In recent years

in vitro

biosynthesis technology has emerged as a groundbreaking alternative

offering a highly versatile

cell-free platform for the efficient synthesis of carbohydrates. This technology integrates multiple enzymes within a single reaction system

enabling them to work synergistically without the constraints of a living cell. This approach offers several significant advantages over traditional cellular synthesis. First

it bypasses the steady-state limitations of cellular systems

allowing for higher reaction rates and enhanced product yields. Second

it provides greater pathway controllability

making it possible to fine-tune reaction conditions for optimal performance. Third

it enhances the atomic economy

as the absence of cellular byproducts leads to more efficient use of substrates. These attributes make

in vitro

biosynthesis a powerful tool for the scalable and sustainable production of a wide range of carbohydrate products. This review provides a comprehensive overview of the core technological advancements in the

in vitro

carbohydrate synthesis. The discovery

engineering and expression of target enzymes are the foundation of the

in vitro

biosynthesis

providing the basic elements for pathway construction. For the pathway design

thermodynamic driving and cofactor recycling strategies ensure sustained catalytic activity and energy efficiency. Furthermor

the modular design of synthesis pathways allows for flexible configuration and rapid optimization of multi-enzyme systems

while synergistic adaptation mechanisms enable the efficient coordination of enzyme functions. This review also highlights representative applications of

in vitro

biosynthesis technology in the production of various monosaccharides

sugar alcohols

oligosaccharides

polysaccharides

and sugar derivatives

demonstrating its advantages in utilizing low-cost resources

achieving atomic economy

and enabling scalable production. Moreover

the potential of artificial intelligence to further enhance

in vitro

carbohydrate biosynthesis is discussed

particularly in the areas of enzyme discovery and modification

pathway optimization

and process parameter prediction. As the convergence of biology with advanced computational tools continues

in vitro

carbohydrate biosynthesis is expected to accelerate its transition from laboratory research to industrial-scale applications

becoming a key enabler of next-generation green biomanufacturing.

关键词

Keywords

references

CAMERON D E , BASHOR C J , COLLINS J J . A brief history of synthetic biology [J ] . Nature Reviews Microbiology , 2014 , 12 ( 5 ): 381 - 390 .

FOLDI J , CONNOLLY J A , TAKANO E , et al . Synthetic biology of natural products engineering: recent advances across the discover-design-build-test-learn cycle [J ] . ACS Synthetic Biology , 2024 , 13 ( 9 ): 2684 - 2692 .

MENG F K , ELLIS T . The second decade of synthetic biology: 2010—2020 [J ] . Nature Communications , 2020 , 11 : 5174 .

VOIGT C A . Synthetic biology 2020—2030: six commercially-available products that are changing our world [J ] . Nature Communications , 2020 , 11 ( 1 ): 6379 .

CLAASSENS N J , BURGENER S , VÖGELI B , et al . A critical comparison of cellular and cell-free bioproduction systems [J ] . Current Opinion in Biotechnology , 2019 , 60 : 221 - 229 .

WU G , YAN Q , JONES J A , et al . Metabolic burden: cornerstones in synthetic biology and metabolic engineering applications [J ] . Trends in Biotechnology , 2016 , 34 ( 8 ): 652 - 664 .

JUNG S W , YEOM J , PARK J S , et al . Recent advances in tuning the expression and regulation of genes for constructing microbial cell factories [J ] . Biotechnology Advances , 2021 , 50 : 107767 .

RADDE N , MORTENSEN G A , BHAT D , et al . Measuring the burden of hundreds of BioBricks defines an evolutionary limit on constructability in synthetic biology [J ] . Nature Communications , 2024 , 15 : 6242 .

MAO J W , ZHANG H Y , CHEN Y , et al . Relieving metabolic burden to improve robustness and bioproduction by industrial microorganisms [J ] . Biotechnology Advances , 2024 , 74 : 108401 .

CHUBUKOV V , MUKHOPADHYAY A , PETZOLD C J , et al . Synthetic and systems biology for microbial production of commodity chemicals [J ] . NPJ Systems Biology and Applications , 2016 , 2 : 16009 .

CORRÊA G G , DA COSTA RIBEIRO LINS M R , SILVA B F , et al . A modular autoinduction device for control of gene expression in Bacillus subtilis [J ] . Metabolic Engineering , 2020 , 61 : 326 - 334 .

BADRI A , WILLIAMS A , AWOFIRANYE A , et al . Complete biosynthesis of a sulfated chondroitin in Escherichia coli [J ] . Nature Communications , 2021 , 12 : 1389 .

ZHANG Y H P J , ZHU Z G , YOU C , et al . In vitro biotransformation (ivBT): definitions, opportunities, and challenges [J ] . Synthetic Biology and Engineering , 2023 , 1 ( 2 ): 10013 .

WEI X L , YANG X , HU C C , et al . ATP-free in vitro biotransformation of starch-derived maltodextrin into poly-3-hydroxybutyrate via acetyl-CoA [J ] . Nature Communications , 2024 , 15 : 3267 .

石婷 , 宋展 , 宋世怡 , 等 . 体外生物转化(ivBT): 生物制造的新前沿 [J ] . 合成生物学 , 2024 , 5 ( 6 ): 1437 - 1460 .

SHI T , SONG Z , SONG S Y , et al . In vitro biotransformation(ivBT): a new frontier of industrial biomanufacturing [J ] . Synthetic Biology Journal , 2024 , 5 ( 6 ): 1437 - 1460 .

ZHANG Y H P . Production of biocommodities and bioelectricity by cell-free synthetic enzymatic pathway biotransformations: challenges and opportunities [J ] . Biotechnology and Bioengineering , 2010 , 105 ( 4 ): 663 - 677 .

QIN Y M , LI Q Z , FAN L , et al . Biomanufacturing by in vitro biotransformation (ivBT) using purified cascade multi-enzymes [J ] . Advances in Biochemical Engineering/Biotechnology , 2023 , 186 : 1 - 27 .

ZHANG P , WANG J Q , DING X W , et al . Exploration of the tolerance ability of a cell-free biosynthesis system to toxic substances [J ] . Applied Biochemistry and Biotechnology , 2019 , 189 ( 4 ): 1096 - 1107 .

YOU C , CHEN H G , MYUNG S , et al . Enzymatic transformation of nonfood biomass to starch [J ] . Proceedings of the National Academy of Sciences of the United States of America , 2013 , 110 ( 18 ): 7182 - 7187 .

DESVAUX M , GUEDON E , PETITDEMANGE H . Cellulose catabolism by Clostridium cellulolyticum growing in batch culture on defined medium [J ] . Applied and Environmental Microbiology , 2000 , 66 ( 6 ): 2461 - 2470 .

PERSSON O , VALADI A , NYSTRÖM T , et al . Metabolic control of the Escherichia coli universal stress protein response through fructose-6-phosphate [J ] . Molecular Microbiology , 2007 , 65 ( 4 ): 968 - 978 .

RICHARDS G R , PATEL M V , LLOYD C R , et al . Depletion of glycolytic intermediates plays a key role in glucose-phosphate stress in Escherichia coli [J ] . Journal of Bacteriology , 2013 , 195 ( 21 ): 4816 - 4825 .

XING M N , ZHANG X Z , HUANG H . Application of metagenomic techniques in mining enzymes from microbial communities for biofuel synthesis [J ] . Biotechnology Advances , 2012 , 30 ( 4 ): 920 - 929 .

HON J , BORKO S , STOURAC J , et al . EnzymeMiner: automated mining of soluble enzymes with diverse structures, catalytic properties and stabilities [J ] . Nucleic Acids Research , 2020 , 48 ( W1 ): W104 - W109 .

JIA B L , HAN X , HYUN K H , et al . Discovery and mining of enzymes from the human gut microbiome [J ] . Trends in Biotechnology , 2022 , 40 ( 2 ): 240 - 254 .

JOHNSON M , ZARETSKAYA I , RAYTSELIS Y , et al . NCBI BLAST: a better web interface [J ] . Nucleic Acids Research , 2008 , 36 ( Web Server issue ): W5 - W9 .

UniProt Consortium . UniProt: a worldwide hub of protein knowledge [J ] . Nucleic Acids Research , 2019 , 47 ( D1 ): D506 - D515 .

BLUM M , CHANG H Y , CHUGURANSKY S , et al . The InterPro protein families and domains database: 20 years on [J ] . Nucleic Acids Research , 2021 , 49 ( D1 ): D344 - D354 .

DUMORNÉ K , CÓRDOVA D C , ASTORGA-ELÓ M , et al . Extremozymes: a potential source for industrial applications [J ] . Journal of Microbiology and Biotechnology , 2017 , 27 ( 4 ): 649 - 659 .

LI L X , LIU X Q , BAI Y G , et al . High-throughput screening techniques for the selection of thermostable enzymes [J ] . Journal of Agricultural and Food Chemistry , 2024 , 72 ( 8 ): 3833 - 3845 .

WANG Y F , LI W W , WANG D D , et al . Screening and characterization of thermostable xylose isomerase from Rhodothermus marinus for erythrose production from one-carbon source [J ] . Enzyme and Microbial Technology , 2025 , 186 : 110607 .

LI Y J , SHI T , HAN P P , et al . Thermodynamics-driven production of value-added D-allulose from inexpensive starch by an in vitro enzymatic synthetic biosystem [J ] . ACS Catalysis , 2021 , 11 ( 9 ): 5088 - 5099 .

GERLT J A , BOUVIER J T , DAVIDSON D B , et al . Enzyme Function Initiative-Enzyme Similarity Tool (EFI-EST): a web tool for generating protein sequence similarity networks [J ] . Biochimica et Biophysica Acta , 2015 , 1854 ( 8 ): 1019 - 1037 .

BURROUGHS A M , ALLEN K N , DUNAWAY-MARIANO D , et al . Evolutionary genomics of the HAD superfamily: understanding the structural adaptations and catalytic diversity in a superfamily of phosphoesterases and allied enzymes [J ] . Journal of Molecular Biology , 2006 , 361 ( 5 ): 1003 - 1034 .

MAK W S , TRAN S , MARCHESCHI R , et al . Integrative genomic mining for enzyme function to enable engineering of a non-natural biosynthetic pathway [J ] . Nature Communications , 2015 , 6 : 10005 .

SONG Y F , DIMAIO F , WANG R Y , et al . High-resolution comparative modeling with RosettaCM [J ] . Structure , 2013 , 21 ( 10 ): 1735 - 1742 .

MILLER B R Ⅲ , MCGEE T D , SWAILS J M , et al . MMPBSA.py: an efficient program for end-state free energy calculations [J ] . Journal of Chemical Theory and Computation , 2012 , 8 ( 9 ): 3314 - 3321 .

BONETTA R , VALENTINO G . Machine learning techniques for protein function prediction [J ] . Proteins , 2020 , 88 ( 3 ): 397 - 413 .

WANG Y H , HAN S X , WANG Y , et al . Artificial intelligence technology assists enzyme prediction and rational design [J ] . Journal of Agricultural and Food Chemistry , 2025 , 73 ( 12 ): 7065 - 7073 .

YU T H , CUI H Y , LI J C , et al . Enzyme function prediction using contrastive learning [J ] . Science , 2023 , 379 ( 6639 ): 1358 - 1363 .

BOULAHROUF K , ALIOUANE S E , CHEHILI H , et al . EZYDeep: a deep learning tool for enzyme function prediction based on sequence information [J ] . The Open Bioinformatics Journal , 2023 , 16 : e187503622306270 .

TAN Q X , XIAO J , CHEN J Y , et al . ifDEEPre: large protein language-based deep learning enables interpretable and fast predictions of enzyme commission numbers [J ] . Briefings in Bioinformatics , 2024 , 25 ( 4 ): bbae225 .

REETZ M T , SUN Z T , QU G . Enzyme engineering: selective catalysts for applications in biotechnology, organic chemistry, and life science [M ] . New York : Wiley-VCH , 2023 .

VICTORINO DA SILVA AMATTO I , GONSALES DA ROSA-GARZON N , ANTÔNIO DE OLIVEIRA SIMÕES F , et al . Enzyme engineering and its industrial applications [J ] . Biotechnology and Applied Biochemistry , 2022 , 69 ( 2 ): 389 - 409 .

MAZURENKO S , PROKOP Z , DAMBORSKY J . Machine learning in enzyme engineering [J ] . ACS Catalysis , 2020 , 10 ( 2 ): 1210 - 1223 .

SONG Z D , ZHANG Q F , WU W H , et al . Rational design of enzyme activity and enantioselectivity [J ] . Frontiers in Bioengineering and Biotechnology , 2023 , 11 : 1129149 .

CHICA R A , DOUCET N , PELLETIER J N . Semi-rational approaches to engineering enzyme activity: combining the benefits of directed evolution and rational design [J ] . Current Opinion in Biotechnology , 2005 , 16 ( 4 ): 378 - 384 .

WATERHOUSE A , BERTONI M , BIENERT S , et al . SWISS-MODEL: homology modelling of protein structures and complexes [J ] . Nucleic Acids Research , 2018 , 46 ( W1 ): W296 - W303 .

ABRAMSON J , ADLER J , DUNGER J , et al . Accurate structure prediction of biomolecular interactions with AlphaFold 3 [J ] . Nature , 2024 , 630 ( 8016 ): 493 - 500 .

SOUSA S F , FERNANDES P A , RAMOS M J . Protein-ligand docking: current status and future challenges [J ] . Proteins , 2006 , 65 ( 1 ): 15 - 26 .

YANG C , CHEN E A , ZHANG Y K . Protein-ligand docking in the machine-learning era [J ] . MOLECULES , 2022 , 27 ( 14 ): 4568 .

VAN DER SPOEL D , LINDAHL E , HESS B , et al . GROMACS: fast, flexible, and free [J ] . Journal of Computational Chemistry , 2005 , 26 ( 16 ): 1701 - 1718 .

LEE T S , ALLEN B K , GIESE T J , et al . Alchemical binding free energy calculations in AMBER20: advances and best practices for drug discovery [J ] . Journal of Chemical Information and Modeling , 2020 , 60 ( 11 ): 5595 - 5623 .

TIAN C Y , YANG J G , LIU C , et al . Engineering substrate specificity of HAD phosphatases and multienzyme systems development for the thermodynamic-driven manufacturing sugars [J ] . Nature Communications , 2022 , 13 : 3582 .

ZHANG T , LIU P , WEI H L , et al . Protein engineering of glucosylglycerol phosphorylase facilitating efficient and highly regio- and stereoselective glycosylation of polyols in a synthetic system [J ] . ACS Catalysis , 2022 , 12 ( 24 ): 15715 - 15727 .

ROMERO P A , ARNOLD F H . Exploring protein fitness landscapes by directed evolution [J ] . Nature Reviews Molecular Cell Biology , 2009 , 10 ( 12 ): 866 - 876 .

COPP J N , HANSON-MANFUL P , ACKERLEY D F , et al . Error-prone PCR and effective generation of gene variant libraries for directed evolution [J ] . Methods in Molecular Biology , 2014 , 1179 : 3 - 22 .

HUANG R , CHEN H , ZHONG C , et al . High-throughput screening of coenzyme preference change of thermophilic 6-phosphogluconate dehydrogenase from NADP + to NAD + [J ] . Scientific Reports , 2016 , 6 : 32644 .

MA C L , LIU M X , YOU C , et al . Engineering a diaphorase via directed evolution for enzymatic biofuel cell application [J ] . Bioresources and Bioprocessing , 2020 , 7 ( 1 ): 23 .

ZHOU W , HUANG R , ZHU Z G , et al . Coevolution of both thermostability and activity of polyphosphate glucokinase from Thermobifida fusca YX [J ] . Applied and Environmental Microbiology , 2018 , 84 ( 16 ): e01224-18 .

ZHU Y M , CHEN P , DONG Q Z , et al . Protein engineering of transaminase facilitating enzyme cascade reaction for the biosynthesis of azasugars [J ] . iScience , 2024 , 27 ( 3 ): 109034 .

YANG K K , WU Z , ARNOLD F H . Machine-learning-guided directed evolution for protein engineering [J ] . Nature Methods , 2019 , 16 ( 8 ): 687 - 694 .

SUMIDA K H , NÚÑEZ-FRANCO R , KALVET I , et al . Improving protein expression, stability, and function with ProteinMPNN [J ] . Journal of the American Chemical Society , 2024 , 146 ( 3 ): 2054 - 2061 .

BIAN J H , TAN P , NIE T , et al . Optimizing enzyme thermostability by combining multiple mutations using protein language model [J ] . mLife , 2024 , 3 ( 4 ): 492 - 504 .

REUTEN R , NIKODEMUS D , OLIVEIRA M B , et al . Maltose-binding protein (MBP), a secretion-enhancing tag for mammalian protein expression systems [J ] . PLoS One , 2016 , 11 ( 3 ): e0152386 .

GUERRERO F , CIRAGAN A , IWAÏ H . Tandem SUMO fusion vectors for improving soluble protein expression and purification [J ] . Protein Expression and Purification , 2015 , 116 : 42 - 49 .

REBAY I , FEHON R G . Preparation of soluble GST fusion proteins [J ] . Cold Spring Harbor Protocols , 2009 , 2009 ( 11 ): pdb.prot4996.

XIE X , WU P , HUANG X C , et al . Retro-protein XXA is a remarkable solubilizing fusion tag for inclusion bodies [J ] . Microbial Cell Factories , 2022 , 21 ( 1 ): 51 .

NGUYEN T K M , KI M R , SON R G , et al . The NT11, a novel fusion tag for enhancing protein expression in Escherichia coli [J ] . Applied Microbiology and Biotechnology , 2019 , 103 ( 5 ): 2205 - 2216 .

DE MARCO A . Protocol for preparing proteins with improved solubility by co-expressing with molecular chaperones in Escherichia coli [J ] . Nature Protocols , 2007 , 2 ( 10 ): 2632 - 2639 .

DING Z D , GUAN F F , XU G S , et al . MPEPE, a predictive approach to improve protein expression in E . coli based on deep learning [J ] . Computational and Structural Biotechnology Journal , 2022 , 20 : 1142 - 1153 .

KHADYE V S , SAWANT S , SHAIKH K , et al . Optimal secretion of thermostable β-glucosidase in Bacillus subtilis by signal peptide optimization [J ] . Protein Expression and Purification , 2021 , 182 : 105843 .

PAN Y J , YANG J , WU J P , et al . Current advances of Pichia pastoris as cell factories for production of recombinant proteins [J ] . Frontiers in Microbiology , 2022 , 13 : 1059777 .

INAN M , ARYASOMAYAJULA D , SINHA J , et al . Enhancement of protein secretion in Pichia pastoris by overex pression of protein disulfide isomerase [J ] . Biotechnology and Bioengineering , 2006 , 93 ( 4 ): 771 - 778 .

张以恒 , 陈雪梅 , 石婷 . 生物制造的市本率(PC值): 定义与应用 [J ] . 合成生物学 , 2025 , 6 ( 1 ): 8 - 17 .

ZHANG Y H , CHEN X M , SHI T . Price to cost-of-raw-materials ratio (PC) of biomanufacturing: definition and application [J ] . Synthetic Biology Journal , 2025 , 6 ( 1 ): 8 - 17 .

BEBER M E , GOLLUB M G , MOZAFFARI D , et al . eQuilibrator 3.0: a database solution for thermodynamic constant estimation [J ] . Nucleic Acids Research , 2022 , 50 ( D1 ): D603 - D609 .

GAO J L , MA S H , MAJOR D T , et al . Mechanisms and free energies of enzymatic reactions [J ] . Chemical Reviews , 2006 , 106 ( 8 ): 3188 - 3209 .

YANG J G , ZHANG T , TIAN C Y , et al . Multi-enzyme systems and recombinant cells for synthesis of valuable saccharides: advances and perspectives [J ] . Biotechnology Advances , 2019 , 37 ( 7 ): 107406 .

WEN L Q , HUANG K , WEI M H , et al . Facile enzymatic synthesis of ketoses [J ] . Angewandte Chemie International Edition , 2015 , 54 ( 43 ): 12654 - 12658 .

ANDEXER J N , RICHTER M . Emerging enzymes for ATP regeneration in biocatalytic processes [J ] . ChemBioChem , 2015 , 16 ( 3 ): 380 - 386 .

RESNICK S M , ZEHNDER A J . In vitro ATP regeneration from polyphosphate and AMP by polyphosphate: AMP phosphotransferase and adenylate kinase from Acinetobacter johnsonii 210A [J ] . Applied and Environmental Microbiology , 2000 , 66 ( 5 ): 2045 - 2051 .

MYUNG S , ROLLIN J , YOU C , et al . In vitro metabolic engineering of hydrogen production at theoretical yield from sucrose [J ] . Metabolic Engineering , 2014 , 24 : 70 - 77 .

YOU C , SHI T , LI Y J , et al . An in vitro synthetic biology platform for the industrial biomanufacturing of myo-inositol from starch [J ] . Biotechnology and Bioengineering , 2017 , 114 ( 8 ): 1855 - 1864 .

ZHONG C , YOU C , WEI P , et al . Thermal cycling cascade biocatalysis of myo-inositol synthesis from sucrose [J ] . ACS Catalysis , 2017 , 7 ( 9 ): 5992 - 5999 .

WANG W , LIU M X , YOU C , et al . ATP-free biosynthesis of a high-energy phosphate metabolite fructose 1,6-diphosphate by in vitro metabolic engineering [J ] . Metabolic Engineering , 2017 , 42 : 168 - 174 .

ZHU Y M , LI H Y , LIU P P , et al . Construction of allitol synthesis pathway by multi-enzyme coexpression in Escherichia coli and its application in allitol production [J ] . Journal of Industrial Microbiology & Biotechnology , 2015 , 42 ( 5 ): 661 - 669 .

SATOH Y , TAJIMA K , TANNAI H , et al . Enzyme-catalyzed poly(3-hydroxybutyrate) synthesis from acetate with CoA recycling and NADPH regeneration in Vitro [J ] . Journal of Bioscience and Bioengineering , 2003 , 95 ( 4 ): 335 - 341 .

RELYEA H A , VAN DER DONK W A . Mechanism and applications of phosphite dehydrogenase [J ] . Bioorganic Chemistry , 2005 , 33 ( 3 ): 171 - 189 .

ZHAO H M , VAN DER DONK W A . Regeneration of cofactors for use in biocatalysis [J ] . Current Opinion in Biotechnology , 2003 , 14 ( 6 ): 583 - 589 .

LI J , MU S C , YANG J G , et al . Glycosyltransferase engineering and multi-glycosylation routes development facilitating synthesis of high-intensity sweetener mogrosides [J ] . iScience , 2022 , 25 ( 10 ): 105222 .

BAULER P , HUBER G , LEYH T , et al . Channeling by proximity: the catalytic advantages of active site colocalization using Brownian dynamics [J ] . The Journal of Physical Chemistry Letters , 2010 , 1 ( 9 ): 1332 - 1335 .

FAN L W , WANG Y , TUYISHIME P , et al . Engineering artificial fusion proteins for enhanced methanol bioconversion [J ] . Chembiochem , 2018 , 19 ( 23 ): 2465 - 2471 .

TIAN J W , JIA R Q , DONG W G , et al . One-step purification and immobilization of recombinant proteins using SpyTag/SpyCatcher chemistry [J ] . Biotechnology Letters , 2021 , 43 ( 5 ): 1075 - 1087 .

STAHL S W , NASH M A , FRIED D B , et al . Single-molecule dissection of the high-affinity cohesion-dockerin complex [J ] . Proceedings of the National Academy of Sciences of the United States of America , 2012 , 109 ( 50 ): 20431 - 20436 .

SUN X M , ZHANG T , LIU Y L , et al . Self-assembled multienzyme complex facilitates synthesis of glucosylglycerol from maltodextrin and glycerol [J ] . Journal of the Science of Food and Agriculture , 2024 , 104 ( 1 ): 266 - 272 .

HAN P P , ZHOU X G , YOU C . Efficient multi-enzymes immobilized on porous microspheres for producing inositol from starch [J ] . Frontiers in Bioengineering and Biotechnology , 2020 , 8 : 380 .

HAN P P , YOU C , LI Y J , et al . High-titer production of myo -inositol by a co-immobilized four-enzyme cocktail in biomimetic mineralized microcapsules [J ] . Chemical Engineering Journal , 2023 , 461 : 141946 .

YANG J G , SONG W , CAI T , et al . De novo artificial synthesis of hexoses from carbon dioxide [J ] . Science Bulletin , 2023 , 68 ( 20 ): 2370 - 2381 .

WANG L , DASH S , NG C Y , et al . A review of computational tools for design and reconstruction of metabolic pathways [J ] . Synthetic and Systems Biotechnology , 2017 , 2 ( 4 ): 243 - 252 .

JANG W D , KIM G B , KIM Y J , et al . Applications of artificial intelligence to enzyme and pathway design for metabolic engineering [J ] . Current Opinion in Biotechnology , 2022 , 73 : 101 - 107 .

WEI F , CAI J Y , MAO Y F , et al . Unveiling metabolic engineering strategies by quantitative heterologous pathway design [J ] . Advanced Science , 2024 , 11 ( 45 ): 2404632 .

IZUMORI K . Izumoring: a strategy for bioproduction of all hexoses [J ] . Journal of Biotechnology , 2006 , 124 ( 4 ): 717 - 722 .

MEN Y , ZHU P , ZHU Y M , et al . The development of low-calorie sugar and functional jujube food using biological transformation and fermentation coupling technology [J ] . Food Science & Nutrition , 2019 , 7 ( 4 ): 1302 - 1310 .

MEN Y , ZHU Y M , ZENG Y , et al . Co-expression of D-glucose isomerase and D-psicose 3-epimerase: development of an efficient one-step production of D-psicose [J ] . Enzyme and Microbial Technology , 2014 , 64-65 : 1 - 5 .

ZHU P , ZENG Y , CHEN P , et al . A one-pot two-enzyme system on the production of high value-added D-allulose from Jerusalem artichoke tubers [J ] . Process Biochemistry , 2020 , 88 : 90 - 96 .

WEI X L , LI Q Z , HU C C , et al . An ATP-free in vitro synthetic enzymatic biosystem facilitating one-pot stoichiometric conversion of starch to mannitol [J ] . Applied Microbiology and Biotechnology , 2021 , 105 ( 5 ): 1913 - 1924 .

YAN J J , CHEN P , ZENG Y , et al . Production of neoagarobiose from agar through a dual-enzyme and two-stage hydrolysis strategy [J ] . International Journal of Biological Macromolecules , 2020 , 160 : 288 - 295 .

ALI M Y , LIAQAT F , KHAZI M I , et al . Utilization of glycosyltransferases as a seamless tool for synthesis and modification of the oligosaccharides - a review [J ] . International Journal of Biological Macromolecules , 2023 , 249 : 125916 .

TIAN C Y , YANG J G , ZENG Y , et al . Biosynthesis of raffinose and stachyose from sucrose via an in vitro multienzyme system [J ] . Applied and Environmental Microbiology , 2019 , 85 ( 2 ): e02306-18 .

SUN S S , WEI X L , ZHOU X G , et al . Construction of an artificial in vitro synthetic enzymatic platform for upgrading low-cost starch to value-added disaccharides [J ] . Journal of Agricultural and Food Chemistry , 2021 , 69 ( 1 ): 302 - 314 .

SUN S S , YOU C . Disaccharide phosphorylases: structure, catalytic mechanisms and directed evolution [J ] . Synthetic and Systems Biotechnology , 2021 , 6 ( 1 ): 23 - 31 .

CAI T , SUN H B , QIAO J , et al . Cell-free chemoenzymatic starch synthesis from carbon dioxide [J ] . Science , 2021 , 373 ( 6562 ): 1523 - 1527 .

XU X X , ZHANG W , YOU C , et al . Biosynthesis of artificial starch and microbial protein from agricultural residue [J ] . Science Bulletin , 2023 , 68 ( 2 ): 214 - 223 .

LI Y , YOU C , ZHANG Y H P J . In vitro biotransformation of high-titer D-glucarate by stepwise-added enzyme cocktails [J ] . Organic Process Research & Development , 2024 , 28 ( 2 ): 478 - 486 .

MENG D D , WEI X L , BAI X , et al . Artificial in vitro synthetic enzymatic biosystem for the one-pot sustainable biomanufacturing of glucosamine from starch and inorganic ammonia [J ] . ACS Catalysis , 2020 , 10 ( 23 ): 13809 - 13819 .

浏览量

下载量

CSCD

文章被引用时，请邮件提醒。

提交

工具集

关联资源

母乳低聚糖生物合成研究进展

卤化酶在生物催化中的应用：机制解析、定向进化和绿色制造的进展

体外多酶组装与生物级联催化：进展与展望

聚酮化合物非天然延伸单元的生物合成与结构改造应用

合成生物学赋能糖科学与糖工程应用