Genetically engineered whole-cell biocatalyst for efficient CO2 capture by cell surface display of carbonic anhydrase from Bacillus cereus GLRT202 on Escherichia coli
IF 3.7 3区 生物学Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY
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引用次数: 0
Abstract
CO2 sequestration is important for reducing greenhouse effects. Carbonic anhydrase (CA) from bacteria has a promising role because it can be modified by genetic techniques and bioengineering. In this study, the CA from B. cereus GLRT202 (Bc-CA) was genetically engineered and anchored on the surface of E. coli by using the N-domain of the ice nucleation protein from P. syringae (INPN). Both surface-displayed and cytosolic Bc-CA yielded high expression levels of CA when induced with 0.5 mM IPTG. It exhibited no adverse influence on the host cell growth. Additionally, surface-displayed Bc-CA enhanced its stability and specificity compared to cytosolic expressed Bc-CA. The CA activity of whole-cell surface-displayed cells was 1.66-fold higher (5.19 U/mL) than that of the cytosolic form. Besides the advantages of higher activity, the whole-cell displaying CA was comparatively stable, with better storage (at 4 ℃) and resting culture stability (at 37 ℃). The whole-cell biocatalyst induced the calcite precipitation, which indicated that the cell facilitated the CO2 capture. XRD, FTIR, and FESEM characterized calcite precipitates thus obtained. This study demonstrates that Bc-CA can be correctly expressed on the E. coli surface through fusion with the INPN. This leads to an effective whole-cell biocatalyst with enhanced stability and specificity of the enzyme for efficient CO2 capture applications.
二氧化碳封存对减少温室效应非常重要。细菌中的碳酸酐酶(CA)可以通过基因技术和生物工程进行改造,因此具有广阔的应用前景。在这项研究中,对来自 GLRT202()的碳酸酐酶进行了基因工程改造,并利用冰核蛋白(INPN)的 N 域将其锚定在 GLRT202()的表面。在0.5 mM IPTG的诱导下,表面和细胞膜上的CA都有很高的表达量。它对宿主细胞的生长没有不良影响。此外,与细胞质表达的 CA 相比,表面表达的 CA 增强了其稳定性和特异性。全细胞表面表达型细胞的 CA 活性(5.19 U/mL)是细胞质型的 1.66 倍。除了活性高的优点外,全细胞表面表达 CA 的稳定性也相对较好,其贮存稳定性(4 ℃)和静置培养稳定性(37 ℃)均较好。全细胞生物催化剂诱导了方解石沉淀,这表明细胞促进了二氧化碳的捕获。XRD、傅立叶变换红外光谱和 FESEM 对由此获得的方解石沉淀进行了表征。这项研究表明,生物催化剂可以通过与 INPN 的融合在表面正确表达。这就产生了一种有效的全细胞生物催化剂,它提高了酶的稳定性和特异性,可用于高效的二氧化碳捕获。
期刊介绍:
The Biochemical Engineering Journal aims to promote progress in the crucial chemical engineering aspects of the development of biological processes associated with everything from raw materials preparation to product recovery relevant to industries as diverse as medical/healthcare, industrial biotechnology, and environmental biotechnology.
The Journal welcomes full length original research papers, short communications, and review papers* in the following research fields:
Biocatalysis (enzyme or microbial) and biotransformations, including immobilized biocatalyst preparation and kinetics
Biosensors and Biodevices including biofabrication and novel fuel cell development
Bioseparations including scale-up and protein refolding/renaturation
Environmental Bioengineering including bioconversion, bioremediation, and microbial fuel cells
Bioreactor Systems including characterization, optimization and scale-up
Bioresources and Biorefinery Engineering including biomass conversion, biofuels, bioenergy, and optimization
Industrial Biotechnology including specialty chemicals, platform chemicals and neutraceuticals
Biomaterials and Tissue Engineering including bioartificial organs, cell encapsulation, and controlled release
Cell Culture Engineering (plant, animal or insect cells) including viral vectors, monoclonal antibodies, recombinant proteins, vaccines, and secondary metabolites
Cell Therapies and Stem Cells including pluripotent, mesenchymal and hematopoietic stem cells; immunotherapies; tissue-specific differentiation; and cryopreservation
Metabolic Engineering, Systems and Synthetic Biology including OMICS, bioinformatics, in silico biology, and metabolic flux analysis
Protein Engineering including enzyme engineering and directed evolution.