Permeabilized whole-cell biocatalyst containing co-expressed two enzymes facilitates the synthesis of maltoheptaose (G7) from starch

Abdalla, Mohammed; Dai, Yiwei; Chen, Jingjing; Hassanin, Hinawi A.M.; Zhang, Tao

doi:10.1016/j.enzmictec.2022.110057

Cited by 2 publications

(2 citation statements)

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“…Enzymatic catalysis, for instance, may suffer from high cost, low stability, and difficulty in recovery 8 . Whole cell is primarily constrained in low transformation efficiency due to inadequate intracellular mass transport and extracellular transmembrane restriction on large substrates 9 , 10 . In addition, in numerous instances, enzymes are not expressible directly in the cells that cooperate functionally with enzymes, necessitating the utilization of extracellular enzymes to achieve cascade reactions with cells 11 – 13 .…”

Section: Introductionmentioning

confidence: 99%

Co-immobilization of whole cells and enzymes by covalent organic framework for biocatalysis process intensification

Zheng,

Tan

et al. 2024

Nat Commun

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Co-immobilization of cells and enzymes is often essential for the cascade biocatalytic processes of industrial-scale feasibility but remains a vast challenge. Herein, we create a facile co-immobilization platform integrating enzymes and cells in covalent organic frameworks (COFs) to realize the highly efficient cascade of inulinase and E. coli for bioconversion of natural products. Enzymes can be uniformly immobilized in the COF armor, which coats on the cell surface to produce cascade biocatalysts with high efficiency, stability and recyclability. Furthermore, this one-pot in situ synthesis process facilitates a gram-scale fabrication of enzyme-cell biocatalysts, which can generate a continuous-flow device conversing inulin to D-allulose, achieving space-time yield of 161.28 g L−1 d−1 and high stability (remaining >90% initial catalytic efficiency after 7 days of continuous reaction). The created platform is applied for various cells (e.g., E. coli, Yeast) and enzymes, demonstrating excellent universality. This study paves a pathway to break the bottleneck of extra- and intracellular catalysis, creates a high-performance and customizable platform for enzyme-cell cascade biomanufacturing, and expands the scope of biocatalysis process intensification.

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Section: Introductionmentioning

confidence: 99%

Co-immobilization of whole cells and enzymes by covalent organic framework for biocatalysis process intensification

Zheng,

Tan

et al. 2024

Nat Commun

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“…Without prior linearization, it is difficult to predict how it will enter the cell 13,14 . Several chemical and physical approaches, including using lysozyme, ethylenediaminetetraacetic acid (EDTA), and heat treatment, have been employed to enhance membrane permeability 12,15–17 . However, the enzymatic activity of recombinant CDase and MTSase was reduced by 70% and 80% when incubated at 50 °C or above, respectively 4 .…”

Section: Introductionmentioning

confidence: 99%

Permeabilized whole cells containing co‐expressed cyclomaltodextrinase and maltooligosyltrehalose synthase facilitate the synthesis of nonreducing maltoheptaose (N‐G7) from β‐cyclodextrin

et al. 2023

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BACKGROUNDMaltodextrin is an important bulk ingredient in food and other industries; however, drawbacks such as uneven polymerization and high reducibility limit its utilization. Nonreducing maltoheptaose (N‐G7) is a good substitute for maltodextrin owing to its single degree of polymerization and its nonreducing properties. In this study, in vitro cell factory biotransformation of β‐cyclodextrin (β‐CD) to N‐G7 is demonstrated using coexpressed cyclomaltodextrinase (CDase, EC 3.2.1.54) and maltooligosyltrehalose synthase (MTSase, EC 5.4.99.15). However, the cell membrane prevents β‐CD from entering the cell owing to its large diameter.RESULTSThe amylase‐deficient permeabilized host ΔycjM‐ΔmalS‐ΔlpxM is utilized for the coexpression of recombinant CDase and MTSase. Deletion of lpxM effectively allows the entry of β‐cyclodextrin into the cell, despite its large diameter, without requiring any relevant cell membrane permeability‐promoting reagent. This results in a 28.44% increase in the efficiency of β‐CD entry into the cell, thus enabling intracellular N‐G7 synthesis without the extracellular secretion of recombinant CDase and MTSase. After reacting for 5.5 h, the highest purity of N‐G7 (65.50%) is obtained. However, hydrolysis decreases the purity of N‐G7 to 49.30%, thus resulting in a conversion rate of 40.16% for N‐G7 when the reaction lasts 6 h. Precise control of reaction time is crucial for obtaining high‐purity N‐G7.CONCLUSIONWhole‐cell catalysis avoids cell fragmentation and facilitates the creation of an eco‐friendly, energy‐efficient biotransformation system; thus, it is a promising approach for N‐G7 synthesis. © 2023 Society of Chemical Industry.

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Permeabilized whole-cell biocatalyst containing co-expressed two enzymes facilitates the synthesis of maltoheptaose (G7) from starch

Cited by 2 publications

References 25 publications

Co-immobilization of whole cells and enzymes by covalent organic framework for biocatalysis process intensification

Co-immobilization of whole cells and enzymes by covalent organic framework for biocatalysis process intensification

Permeabilized whole cells containing co‐expressed cyclomaltodextrinase and maltooligosyltrehalose synthase facilitate the synthesis of nonreducing maltoheptaose (N‐G7) from β‐cyclodextrin

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