Facing the protein supply-demand contradiction brought about by global population growth and the resource and environmental constraints of traditional agricultural production models, developing efficient and sustainable new protein sources has become a major strategic direction for practicing the all-encompassing approach to food, ensuring national food security, and meeting the people's needs for a better life. Microbial proteins, as a new type of biomanufacturing products (microbial protein or specific functional proteins) produced by microbial cell factories, have attracted much attention due to the high production efficiency and strong adaptability to raw materials. This article systematically reviews the latest research progress and industrialization status of microbial protein and functional protein in the food and pharmaceutical fields, analyzes the key technologies from strain creation, process scale-up to product application development, as well as the multiple challenges of cost structure, market regulation, and consumer acceptance faced in the industrialization process. Finally, this article makes an outlook on the prospects of microbial protein in high-value fields such as feed replacement, food innovation, pharmaceuticals, and cosmetics. It aims to provide a systematic reference for technological innovation, industrial layout, and policy formulation in China's microbial protein industry and to assist in the high-quality development of the industry.
Dipeptidyl peptidase 4(DPP4)is a membrane-anchored protein ubiquitously expressed across various cell types,primarily responsible for cleaving N-terminal X-Pro or X-Ala dipeptides from polypeptides.It plays a crucial role in glucose metabolism by degrading incretin hormones,thereby regulating blood glucose homeostasis and establishing itself as a key therapeutic target for type 2 diabetes mellitus.In addition,DPP4 has emerged as a potential biomarker and intervention target in pathologies such as fibrosis,inflammation,and cancer.In terms of therapeutic strategies,in addition to conventional small-molecule inhibitors,DPP4-inhibitory peptides have shown significant efficacy in vitro,underscoring their potential for the control of high blood glucose.Advancements in the development of DPP4-specific antibodies are creating new opportunities for diagnosing and treating DPP4-related diseases.This review systematically delineates the established enzymatic functions and blood glucose-rising mechanisms of DPP4,its novel pathological roles in fibrotic,inflammatory,and oncological processes,and discusses recent progress in diverse intervention strategies,including small molecules,inhibitory peptides,and antibodies.It helps deepen the overall understanding of the multifaceted biological functions of DPP4 and its associations with disease,and provides a theoretical basis for optimizing DPP4-targeted therapeutic strategies and developing novel treatment approaches.
Glutaric acid is an important monomer for the synthesis of polymeric materials such as polyesters and polyamides, whereas its bio-based production efficiency has long been limited. In this study, we focused on the key metabolic bottlenecks in glutaric acid biosynthesis and constructed a high-producing Corynebacterium glutamicum cell factory. First, the glutaric acid biosynthetic pathway was integrated into the genome of a high-lysine-producing C. glutamicum strain to establish a basal production strain. Subsequently, modular metabolic engineering strategies were employed to strengthen the biosynthetic modules and eliminate byproduct formation pathways, resulting in a glutaric acid titer of 78.67 g/L. To alleviate α-ketoglutarate depletion, a cofactor regeneration cycle composed of l-glutamate oxidase and catalase was introduced, further increasing the glutaric acid titer to 84.43 g/L. Finally, pH and temperature were optimized in a 5 L bioreactor, leading to a final glutaric acid titer of 97.63 g/L with a yield of 0.36 g/g. This study lays a foundation for the construction of high-producing glutaric acid cell factories based on C. glutamicum.
We assessed the effects of terminal moist-heat sterilization on the structure of poly(3-hydroxybutyrate-co-4-hydroxybutyrate)(P34HB)microspheres and the composite hydrogels with hyaluronic acid(HA),aiming to define an acceptable process window.According to pharmacopeial and sterilization standards,we selected 121℃as the sterilization temperature and set holding time of 10,20,and 30 min,with non-sterilized samples as controls.P34HB microspheres were first blended with HA and then subjected to moist-heat sterilization(co-sterilization).Gel permeation chromatography(GPC),differential scanning calorimetry(DSC),Fourier transform infrared spectroscopy(FTIR),particle size analysis,and scanning electron microscopy(SEM)were employed to characterize the changes in molecular weight,thermal properties,chemical structure,and morphology of microspheres.The injectability and stability of the composite hydrogels were assessed based on oscillatory rheology,extrusion force,cohesiveness,and centrifugation stability.In addition,bioburden and sterility tests were carried out.After sterilization at 121℃for 10-20 min,P34HB microspheres showed a slight decrease in molecular weight and basically unchanged melting temperature,particle size,and morphology,and the composite hydrogels retained suitable viscoelasticity and a steady extrusion force of 12-15 N.After sterilization for 30 min,the microspheres showed a significantly decreased molecular weight and enhanced cold crystallization and double melting peaks in DSC,and the composite hydrogels presented increased fluctuations in elastic modulus and extrusion force as well as signs of phase separation.All sterilized groups passed sterility testing.We conclude that moist-heat sterilization at 121℃for 10-20 min is effective while retaining the complete structure of P34HB microspheres and good injectability of the composite hydrogels,and it is thus suitable as the condition for terminal moist-heat sterilization.Compared with literature data for polycaprolactone(PCL)and poly(L-lactide)(PLLA),P34HB shows superior tolerance to moist-heat and appears to be a promising material for prefilled ready-to-use soft tissue filler injections.The findings provide practical guidance for formulation and process design.
Gelsemium elegans(Gardner and Champ.)Benth.,a highly toxic plant of Loganiaceae,contains monoterpenoid indole alkaloids(MIAs)as its primary bioactive components.Although these alkaloids exhibit significant toxicity,they also exhibit diverse pharmacological activities.The biosynthesis of G.elegans alkaloids originates from the terpenoid pathway and tryptamine pathway,converging at the pivotal intermediate strictosidine.However,the modification reactions in its downstream pathway and the associated key enzyme genes remain to be systematically elucidated.In terms of extraction and separation,techniques such as microwave-assisted extraction demonstrates improved efficiency,and the combination of high-speed countercurrent chromatography with preparative liquid chromatography facilitates the isolation of high-purity monomeric compounds.Pharmacological studies have shown that G.elegans alkaloids possess multiple biological activities,including antitumor,anti-inflammatory,analgesic,anxiolytic,immunomodulatory,cardiovascular protective,mydriatic,and growth-promoting effects,with compounds such as koumine demonstrating particularly prominent activities.Nevertheless,challenges such as the complex alkaloid biosynthetic pathway,difficulties in extraction and purification,and unclear toxicity mechanisms severely hinder their clinical application and translation.This review systematically summarizes the research progress in the synthesis,extraction,and pharmacological effects of alkaloids from G.elegans,aiming to provide reference for the in-depth development and safe utilization of G.elegans resources.
Recombinant elastomeric protein exhibits excellent biocompatibility, tunable mechanical properties, and biomimetic microenvironment characteristics, offering broad application prospects in the biomedical and food sectors. However, its low production efficiency has hindered further research and industrial translation. To address this challenge, this study established a stable and efficient large-scale preparation process through a multi-stage optimization strategy. First, codon optimization of ure(80) increased the yield to 0.42 g/L. Targeted truncation analysis identified amino acid residues 50-59 as a key region limiting expression. Subsequent secondary codon optimization and mRNA structure optimization targeting this region, combined with rational design of the ribosome-binding site (RBS), led to the construction of a high-producing engineered strain, Escherichia coli BL21(DE3)/pET-28a-80opt-R1, which achieved a yield of 0.62 g/L-a two-fold increase compared with the initial yield. Second, response surface methodology was employed to optimize the concentrations of carbon source, nitrogen source, trace elements, and vitamin B1 in a minimal salt medium, while fermentation parameters including induction temperature, induction duration, pH, and IPTG concentration were optimized, increasing the protein yield to 3.5 g/L. Upon scale-up in a fermenter, leveraging its superior mass transfer and process control capabilities, the final yield reached 5.0 g/L, representing a 16.7-fold increase over the initial yield (0.3 g/L). Cellular functional assays confirmed that cell proliferation and adhesion were not adversely affected. This study provides a reliable process for the large-scale preparation of recombinant elastomeric protein and offers a reference for the industrial production of other complex proteins.
Stay-green(sgr)genes encode magnesium dechelatases,which catalyze chlorophyll degradation and play a pivotal role in regulating plant senescence and determining crop yields.This study systematically investigated the biological functions of Sbsgr genes in sorghum and their roles in stress responses,aiming to provide a reference for further research on the molecular mechanisms of sgr genes.Through integrated bioinformatics and molecular biological approaches,we systematically characterized the Sbsgr gene family in sorghum in terms of gene structures,protein physicochemical properties,promoter cis-acting elements,expression profiles,interacting proteins,and natural allelic variations.The results indicated that sorghum carried two sgr homologous genes.Sbsgr1 was an sgr homolog,while Sbsgr2 was an sgr-like homolog.The promoter region of Sbsgr harbored cis-acting elements associated with drought stress and abscisic acid responses.The expression patterns of the two genes differed significantly.Sbsgr1 was primarily expressed in leaves,panicles,and seeds,whereas Sbsgr2 showed high expression levels in leaves.Under drought,low nitrogen,and infection by Sporisorium reilianum,the expression of Sbsgr1 was significantly upregulated,while that of Sbsgr2 was suppressed.Three varieties with strong stay-green traits were preliminarily identified,and two SNP sites in Sbsgr1 were detected in stay-green lines such as'B35'.The results of this study can provide a reference for further in-depth research on the molecular mechanism of the sgr gene.
To address the antigen display limitations of current vaccine carriers, we engineered the T4 bacteriophage into a high-capacity platform. The T4 bacteriophage has advantages such as structural stability, high loading capacity, and easy production. However, the presence of high-copy endogenous Soc protein on its capsid surface severely restricts the effective display of exogenous proteins. To overcome the aforementioned spatial limitations, we employed the CRISPR/Cas9 system to precisely knockout the Soc gene of T4 bacteriophage. We successfully achieved the knockout of the Soc gene by co-transferring the three plasmid systems-pCas, pTargetF-sgRNA, and pMD19-T-Soc-arm-into Escherichiacoli TG1, inducing the expression of Cas9 with L-arabinose, and then infecting the engineered bacteria with the wild-type T4 bacteriophage (named T4 WT). The PCR, SDS-PAGE, and sequencing results confirmed a Soc gene-deficient T4 bacteriophage mutant strain (named T4ΔSoc) was successfully constructed. This mutant strain had comparable growth, thermal stability, and pH stability to the wild type, and maintained complete infectivity. After continuous passage for five generations, it remained stable in terms of genome, protein composition, and phage plaque phenotype, with no occurrence of revertant mutations. In conclusion, T4ΔSoc is stable both genetically and phenotypically, effectively breaking through the limitation of the restricted display space of T4 WT. This study provides a reliable vector tool and technical foundation for constructing an efficient and stable antigen display and vaccine delivery platform and has good application potential in vaccine research and the construction of targeted delivery systems.
DNA methylation is an important epigenetic modification that plays a significant role in various processes of plant growth and development,particularly in flower development.Studies on multiple horticultural plants have revealed that during flower development,changes in environmental temperature,photoperiod,and endogenous phytohormones can lead to either a decrease or an increase in DNA methylation.This process primarily regulates phenotypes such as flower bud differentiation,flower sex differentiation,flowering time,petal color,petal number,and flower senescence by modulating the expression of flower development-related genes(e.g.,FT,FLC,AP1,and MYB).Treatment with DNA methylation inhibitors during flower development can reduce the level of DNA methylation,thereby altering flowering time,flower bud differentiation processes,and related flowering phenotypes.This article first provides an overview of the regulatory mechanisms of DNA methylation and its effects on plant growth and development.It then reviews the types and mechanisms of DNA methylation inhibitors,with a focus on the research progress in DNA methylation regulation of flower bud differentiation,flower sex differentiation,flowering process,and flowering phenotypes.The findings offer insights for further unveiling the molecular roles of DNA methylation in plant flower development and its applications in agricultural production.
The Katanin complex is the only protein complex identified in plants to date that possesses microtubule-severing activity,playing a pivotal role in cell shape remodeling and growth.This complex consists of a catalytic subunit KTN1 and a regulatory subunit KTN80.In Arabidopsis thaliana,KTN80 is encoded by four homologous genes,though their specific functions remain unclear.To elucidate the role of KTN80 in root morphogenesis,we performed a detailed expression analysis of KTN80s(KTN80.1,KTN80.2,KTN80.3,and KTN80.4)via β-glucuronidase histochemical staining.Our findings confirmed that all the four genes were actively expressed in roots,yet they exhibited distinct spatial expression patterns.The phenotypic analysis showed that the ktn80.1234 mutant exhibited abnormal root development,characterized by shortened and robust primary root but significantly extended root hairs compared to the wild type.Microscopic examination showed marked reductions in both the meristematic zone length and the elongation zone length,accompanied by enlarged cells exhibiting isotropic expansion in the mutant.To probe the underlying cellular cause,we employed live-cell imaging to observe cortical microtubule dynamics and discovered disorganized microtubule arrays in the mutant,which impeded normal,rapid cell elongation.Further investigation of the root meristem showed that in ktn80.1234 cells,both the preprophase band(PPB)and the phragmoplast had abnormal microtubule arrays.The PPB microtubules were disorganized,while some phragmoplast microtubules displayed abnormal oblique arrangements.In conclusion,our results demonstrate that KTN80 is essential for root morphogenesis by precisely regulating the reorganization of the cellular microtubule cytoskeleton.This study provides new insights for understanding the role of the Katanin complex in plant growth and development.
RNA interference(RNAi)is a specific post-transcriptional gene silencing mechanism in eukaryotes that mediates homologous mRNA degradation based on double-stranded RNA(dsRNA).Against the backdrop of exploring and innovating green bio-agriculture,the development of environmentally friendly and reliable biopesticides has become a new trend in plant disease control.At present,RNAi-based biopesticides,as a new type of pesticides,have become a current research hotspot and a priority of development due to their strong specificity and environmental friendliness that distinguish them from chemical pesticides.This paper reviews the biosynthesis system of dsRNA and various drug delivery routes for maintaining the stability of dsRNA,with the aim of providing references for domestic research in related fields.
Komagataella phaffii is widely recognized as a premier host for the production of recombinant proteins and value-added metabolites, owing to its low background secretion of endogenous proteins, strong capacity for heterologous protein secretion, and robust growth and metabolic performance under industrially relevant fermentation conditions. In recent years, rapid progress in genome editing technologies and synthetic biology toolkits has markedly improved the precision and efficiency of gene function interrogation, metabolic pathway reconstruction, and dynamic regulation in K. phaffii, thereby continuously strengthening its performance as a microbial cell factory. Consequently, beyond its established roles in producing recombinant proteins, industrial enzymes, and vaccine antigens, K. phaffii has also demonstrated substantial potential for the biosynthesis of natural products, biopharmaceutical molecules, and emerging biomaterials. This review systematically summarizes the evolution of genome engineering technologies in K. phaffii, spanning the transition from conventional recombination-based methods to next-generation precision editing tools. We highlight recent advances, optimization strategies, and engineering practices of CRISPR/Cas and related systems in this host. Moreover, in light of emerging research trends, we discuss key challenges and opportunities associated with improving editing efficiency, enabling rapid assembly of complex metabolic pathways, and accelerating industrial translation, thereby providing a reference for the rational engineering of Komagataella phaffii and its broader applications in synthetic biology and biomanufacturing.
Nicotinamide riboside (NR), an essential precursor for nicotinamide adenine dinucleotide (NAD+), has garnered increasing attention for its anti-aging properties and diverse pharmacological activities. Developing an efficient, sustainable, and environmentally friendly synthesis approach is of great significance for the industrial production of NR. In this study, we developed a modular metabolic engineering strategy to construct an engineered strain capable of synthesizing NR from nicotinamide efficiently. First, six genes, including rihA, were deleted to block the degradation pathways of NR and its key intermediates. Second, the key enzyme BaPrsL135I and the NR transporter MdtL were introduced, significantly enhancing NR accumulation and extracellular export. Subsequently, pncA was deleted to improve the nicotinamide utilization efficiency. Furthermore, precursor supply was substantially increased by relieving purR-mediated feedback regulation and strengthening the pentose phosphate pathway. Through the coordinated optimization of these modules and introduction of the toxin-antitoxin system, the engineered strain NK22 produced 3.84 g/L NR after 36 h of shake-flask fermentation. Moreover, the strain achieved a final NR titer of 27.23 g/L in a 5 L fed-batch bioreactor under antibiotic-free conditions. This study establishes a next-generation NR-producing strain capable of antibiotic-free fermentation and provides a robust framework for the development of efficient and sustainable microbial cell factories for NR and its high-value derivatives.
This study developed kinetic models to describe the growth and metabolism of MDCK adherent cells cultured in basket bioreactors. These models were used to predict cell density, nutrient consumption, and metabolite concentrations at key stages of the culture process in basket bioreactors, helping to overcome sampling limitations caused by environmental conditions and bioreactor design. Experimental data, including cell density and glucose and lactate levels were obtained at different time points from 5 L bioreactors. Model parameters were estimated by nonlinear fitting in MATLAB. Kinetic models were established based on the Logistic and Luedeking-Piret equations, and a perfusion dilution item was introduced considering the perfusion culture. The prediction performance of the models was evaluated based on the goodness-of-fit, regression, and error distribution. The models were then validated with independent experimental data from 40 L basket bioreactors. The results showed that MDCK cells cultured in the 5 L bioreactor entered the plateau phase between 96 h and 108 h, with the maximum cell density reaching (795.13±16.22)×104 cells/mL. The specific growth rate peaked with a value of 1.01/d at the time point of 36 h. During the plateau phase, the maximum lactate production rate was 1.71 mmol/(L·h), while the glucose consumption rate reached 2.21 mmol/(L·h). The kinetic models derived from the 5 L bioreactor data showed good agreement with experimental results, with R2 values exceeding 0.95. When being applied to validation data from three independent 40 L bioreactor batches, the models consistently achieved R2 values above 0.95. Offline cell densities measured by digestion at 96 h and 120 h showed no significant differences from the model predictions (P=0.48, P=0.92). The results indicated that the kinetic models developed with 5 L bioreactor data could accurately predict the growth and metabolic behavior of MDCK cells in the 40 L system. In addition, the results demonstrated that the operational performance of the 40 L bioreactor was comparable to that of the 5 L system. This confirmed the reliability of the established models for scale-up applications. By quantifying key parameters, including the specific growth rate, glucose consumption rate, and lactate production rate, the models provide a sound theoretical basis for optimizing basket bioreactor processes. Moreover, they offer strong technical support for improving production efficiency and quality control in industrial-scale manufacturing.
Microalgae represent a sustainable, green, and efficient biomass resource with enormous potential applications in food, energy, environment, and health industries. To increase the yield per unit area and the overall efficiency of production, selecting suitable locations for production facilities is critically important. The hot-dry valley regions of southwest China are characterized by long sunshine duration, high temperatures, a dry climate, scarce flat land, predominantly mountainous terrain, and poor transportation accessibility. These natural and socioeconomic constraints have severely limited the development of conventional agriculture. However, these same conditions confer unique and highly favorable advantages for the development of a microalgal industry. This review comprehensively analyzes the advantages of developing a microalgal industry in hot-dry valley regions from the perspectives of climate, geography, and socioeconomic conditions. It further elaborates on the promising prospects of microalgal biotechnology in key areas such as the production of carotenoids, the treatment of animal farming wastewater, and the manufacture of future human food products. The primary objective is to advocate research, promotion, and application of microalgal biotechnology in China's hot-dry valley regions, thereby accelerating the upgrading and high-quality development of the national microalgal industry and contributing to sustainable development goals.
Polyethylene terephthalate (PET) is one of the most widely produced synthetic plastics globally, posing serious environmental challenges due to its resistance to natural degradation. Enzymatic degradation offers a sustainable solution for PET recycling. However, natural PET hydrolases often suffer from limited catalytic efficiency, and existing screening methods are labor-intensive with low throughput. To overcome the limitations of conventional methods in throughput and efficiency, this study developed a fluorescent nanoprobe technology combining substrate authenticity with high detection sensitivity, which was integrated into an ultra-high-throughput fluorescence-activated droplet sorting (FADS) platform for the single-cell screening and directed evolution of PET hydrolases. Fluorescein dilaurate (FDL)-loaded PET nanoparticles (PET-FDL NPs) were synthesized as specific fluorogenic probes. By combining Escherichia coli surface display of mScarletI-leaf-branch compost cutinase (LCC) fusion proteins with a dual-fluorescence ratiometric assay (fluorescein/mScarletI), we achieved precise normalization of enzymatic activity against expression variations. An LCC mutant library generated by error-prone PCR (epPCR) was screened by FADS at a throughput of 107 droplets per day. The results showed that FADS enrichment significantly increased the proportion of positive droplets. Subsequent microplate rescreening revealed that this strategy improved the positive hit rate from ~5% (plate screening) to 44%, establishing a robust and scalable workflow for directed evolution of PET hydrolases.
This study aimed to investigate the role of protein tyrosine kinase-like 7(PTK7)in the progression of fibrosis of renal tubular epithelial cells and to elucidate its underlying molecular mechanisms,thereby evaluating the potential of PTK7 as a therapeutic target for renal fibrosis.Immunohistochemistry,Western blotting,and quantitative real-time polymerase chain reaction(qRT-PCR)were performed to examine PTK7 expression in a mouse model of renal fibrosis.Public single-cell transcriptomic datasets were employed to analyze the expression distribution of PTK7 among different renal cell types.A stable PTK7-knockout transformed C3H mouse kidney-1(TCMK-1)cell line was generated via clustered regularly interspaced short palindromic repeats/CRISPR-associated protein 9(CRISPR/Cas9),and following transforming growth factor-beta 1(TGF-β1)stimulation,cellular morphological changes were observed.The expression levels of epithelial-mesenchymal transition(EMT)-related proteins(E-cadherin and vimentin)and fibrosis-related proteins(α-SMA,fibronectin,and collagen I)were assessed.Cell proliferative activity and migratory capacity were evaluated by cell counting kit-8(CCK-8)and wound healing assays,respectively,and the phosphorylation levels of Smad family member 2(Smad2)/Smad family member 3(Smad3)were analyzed to explore the potential mechanisms.PTK7 expression was significantly upregulated in the kidney tissue of the mouse model and was highly expressed in renal tubular epithelial cells.PTK7 knockout suppressed TGF-β1-induced EMT progression and the expression of fibrosis-related proteins(P<0.001),and it attenuated the migratory ability of renal tubular epithelial cells under TGF-β1 stimulation.From a mechanism perspective,PTK7 deficiency effectively inhibited TGF-β1-triggered activation of Smad2/Smad3 phosphorylation.PTK7 promotes the fibrosis of renal tubular epithelial cells through activation of the TGF-β1/Smad signaling pathway,and PTK7 knockout can effectively alleviate fibrosis-related phenotypes at the cellular level.This study provides experimental evidence for further exploration of PTK7 as a potential therapeutic target against renal fibrosis.
We have previously showed that loss of clathrin light chain(CLC)2 and CLC3 in Arabidopsis simultaneously impairs autophagy pathway,thereby resulting in enhanced disease resistance.To understand whether clathrin heavy chain(CHC)proteins play similar roles as CLCs,we used combined approaches of genetics,cell biology,and resistance assays to investigate the roles of CHCs in the autophagy pathway and in autophagy-related immunity.The results showed that the loss of either CHC1 or CHC2 accelerated dark-induced senescence,a hallmark phenotype of defective autophagy,suggesting that the autophagy pathway may be impaired.Consistent with this phenotype,the number of autophagosomes induced under dark treatment conditions was significantly reduced in both chc1-2 and chc2-2 mutants,which indicated that the autophagic flux was significantly reduced in both mutants.The disease resistance assay showed that both mutants exhibited enhanced resistance to the fungal pathogen Golovinomyces cichoracearum,which was highly correlated with elevated reactive oxygen species(ROS)accumulation and increased callose deposition.Together,these findings reveal that CHC-dependent functions are required for efficient autophagy and disruption of clathrin-mediated trafficking leads to the activation of plant immune responses due to an impaired autophagy pathway.This study provides a theoretical basis to enhance disease resistance in crops through manipulation of autophagy pathway.
This issue features a perspectives article grounded in 10 000-ton scale industrial practice, proposing the "neo-chemical industry" as a new lens for understanding biomanufacturing. This commentary discusses why this perspective deserves attention, what new understanding it offers, and what questions it raises for further academic discussion.
Due to its high selectivity, mild reaction conditions, and environmentally friendly characteristics, biocatalysis has shown important potential in the fields of fine chemical synthesis, bioenergy conversion, and environmental governance. However, biocatalysts, including enzymes and microorganisms, generally have problems such as insufficient stability, easy inactivation, difficult recycling, and poor reusability in practical applications, which seriously limit their industrialization process. Recently, metal-organic frameworks (MOFs) have gradually become ideal carriers for constructing efficient biocatalytic systems due to their highly ordered pore structure, ultra-high specific surface area, and designable chemical composition. MOFs can not only achieve efficient immobilization of biocatalysts but also significantly improve catalytic performance through confinement effects, microenvironment regulation, and synergistic catalysis. This study systematically reviews the latest research progress of MOFs in the field of biocatalysis, focusing on the structural advantages, biocatalyst compounding strategies, influencing mechanisms of confinement effects on catalytic performance, and typical applications in biosensing, chiral synthesis, biofuels, and environmental remediation. Finally, it makes an outlook on the current challenges and future development directions of MOFs. This review aims to provide a systematic theoretical foundation and research framework for the rational design and performance optimization of MOF-biocatalyst hybrid systems, and to offer guidance for their engineering applications in green manufacturing and sustainable bioprocesses.
