Critical Reviews in Biotechnology最新文献

{"title":"Revisiting enzyme engineering strategies and reaction mechanisms of lytic polysaccharide monooxygenases (LPMOs).","authors":"Musaddique Hossain, Shalini Das, Subba Reddy Dodda, Kamalpreet Singh, Tiyasa Bhuniya, Tiasha Ghosh, Sudit S Mukhopadhyay","doi":"10.1080/07388551.2025.2612611","DOIUrl":"10.1080/07388551.2025.2612611","url":null,"abstract":"<p><p>Lytic polysaccharide monooxygenases (LPMOs) are auxiliary metalloenzymes that play a crucial role in the degradation of polysaccharides through an oxidative mechanism, distinguishing them from the traditional glycoside hydrolases. Although LPMOs were first identified in 1992, their functional identity and unique oxidative activity were not fully understood until the year 2010. These enzymes cleave at the C1 or C4 position of glycosidic bonds in polysaccharides using molecular oxygen and reductants such as ascorbic acid or cellobiose dehydrogenase (CDH). LPMOs exhibit significant sequence diversity across eight known families and operate via complex mechanisms. Structurally, LPMOs have a conserved active site with a copper ion coordinated by two histidine residues, known as the \"histidine brace\" which is crucial for their oxidative activity Their ability to enhance the efficiency of cellulase enzymes makes them highly valuable in the bio refinery industry. This review focuses on details of: regioselectivity, reaction mechanism, protein engineering strategies, and industrial applications of LPMO. It also emphasizes the building correlation between challenges at the industrial level and their possible solutions through enzyme engineering.</p>","PeriodicalId":10752,"journal":{"name":"Critical Reviews in Biotechnology","volume":" ","pages":"460-476"},"PeriodicalIF":8.7,"publicationDate":"2026-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146104369","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"工程技术","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Brassinosteroid-modified crops: key players for a modern Green Revolution.","authors":"Sunny Ahmar, Marcin Rapacz, Fahad Masoud Wattoo, Ewa Pociecha","doi":"10.1080/07388551.2025.2610339","DOIUrl":"10.1080/07388551.2025.2610339","url":null,"abstract":"<p><p>With the global population projected to reach 9.8 billion by 2050, coupled with extreme weather events and resource scarcity, there is an urgent need to redesign existing Green Revolution varieties (GRVs) for a modern revolution in agriculture. Brassinosteroids (BRs) play crucial roles in shaping plant architecture and hold significant potential for developing modern GRVs that are high-yielding, climate-resilient and environmentally sustainable. Recent advances in crop improvement, particularly by manipulating BR signaling and biosynthesis genes, have already generated semi-dwarf GRVs. These BR-modified GRVs exhibited enhanced overall crop performance, including yield, photosynthetic capacity and nitrogen-use efficiency. However, they still face challenges regarding tolerance to various biotic and abiotic stresses, although studies on BR mutants have indicated that BRs can confer resilience to biotic and abiotic stresses. Despite such studies having made significant progress in exploring the role of BRs, their functions in crop species are not comprehensively understood. Therefore, additional BR mutants need to be developed to make existing crops climate-resilient and to tailor them to specific regional needs, such as increased frost tolerance in Eastern Europe. Ongoing efforts are focused on identifying candidate BR-related genes in crops to reveal the regulatory mechanisms underlying BR signaling and biosynthesis, thereby improving overall performance. This review highlights recent advances in BR research related to crop improvement and stress adaptation. Moreover, it emphasizes the potential of BRs to drive a new generation of BR-modified GRVs and outlines strategies to achieve this by manipulating BR signaling and biosynthesis pathways to develop high-yield, climate-resilient crops.</p>","PeriodicalId":10752,"journal":{"name":"Critical Reviews in Biotechnology","volume":" ","pages":"399-418"},"PeriodicalIF":8.7,"publicationDate":"2026-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147497755","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"工程技术","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"<i>N</i>-Glycosylation differences between recombinant and wildtype glycoproteins revealed by mass spectrometry.","authors":"Yang Zou, Dario Cramer, Albert J R Heck, Karli R Reiding","doi":"10.1080/07388551.2025.2584692","DOIUrl":"10.1080/07388551.2025.2584692","url":null,"abstract":"<p><p>Mass spectrometry (MS) has emerged as a powerful technique to study protein glycosylation. MS on intact denatured or native proteins can reveal all-inclusive glycoproteoform profiles while top-down, middle-down and/or bottom-up MS can uncover the characteristics of individual glycosylation sites. Alternatively, analyzing enzymatically released <i>N</i>-glycans can reveal intricate details on glycan isomers and generate high-throughput data on larger cohorts. All these methods are increasingly applied for the study of both individual glycoproteins and complex glycoprotein mixtures such as those originating from blood plasma or cell lysates. This has increased our knowledge about the complexity of protein glycosylation, but also revealed its huge diversity, which depends not only on the protein but also on the cell-dependent glycosylation machinery that may change with physiological conditions. Currently, multiple glycoproteins are recombinantly produced, for therapeutic applications as well as in the food sector, in host cells of diverse origin, most commonly: <i>E. coli</i> bacteria, yeast cells, insect cells, mammalian CHO or human HEK293 cells. Although glycoproteins of interest might show similar yields when produced in different host cells, an important question remains whether the host cell will or can provide similar or alike glycoproteoform profiles. In this review, we focus on the application of MS-based technologies to study glycosylation profiles of endogenous human glycoproteins and their recombinantly produced counterparts in different host cells. We will discuss in which ways recombinant glycoproteins can differ from their endogenous variants, and the functional consequences.</p>","PeriodicalId":10752,"journal":{"name":"Critical Reviews in Biotechnology","volume":" ","pages":"357-379"},"PeriodicalIF":8.7,"publicationDate":"2026-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147497798","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"工程技术","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Functional genomics in sugarcane breeding: key challenges and strategies.","authors":"Rubab Shabbir, Talha Javed, Sheng-Ren Sun, Zhu-Qing Wang, Wei Zhang, San-Ji Gao, Qin-Nan Wang","doi":"10.1080/07388551.2026.2614075","DOIUrl":"10.1080/07388551.2026.2614075","url":null,"abstract":"<p><p>Sugarcane, a leading source of sugar and bio-energy around the globe stands at the cross-road of genome complexity and agricultural innovation, offering the immense potential to fuel a sustainable future. Functional genomics with its precise identification and manipulation of genes could enable researchers unlock this potential and accelerate the breeding efforts. However, the polyploid genome of sugarcane with: high heterozygosity, high-repetitive DNA content, multiple copies of homo(eo)logous gene, epistatic interaction of alleles, etc., challenges the gene annotation, genome sequencing, genome editing, and phenotypic characterization. Similarly long breeding cycle, low transformation efficiency, time-consuming, and labor-intensive transformation methods further complicates the genome editing. Recent advances of functional genomics are transforming this scenario, such as current availability of reference genome \"R570,\" has provided a significant insight of genome architect and function. Genome wide association studies (GWAS)/genome selection (GS) are enhancing trait-mapping and prediction of breeding values to accelerate the breeding cycles. The current era of smart breeding with integrative bio-informatics, advance genome editing tools, i.e., CRISPR/Cas-systems (Cas-proteins, Cas-RNPs, d-Cas-RNPs, and CRISPRa/i), and high-throughput phenomics offers a significant approach to: overcome transformation bottlenecks, explore complex trait architect and address polyploidy challenges. Therefore, this review summarizes the key challenges and focuses on elaborating recent advances and suggests optimized strategies for future improvement in functional genomics of sugarcane breeding.</p>","PeriodicalId":10752,"journal":{"name":"Critical Reviews in Biotechnology","volume":" ","pages":"419-439"},"PeriodicalIF":8.7,"publicationDate":"2026-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146060691","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"工程技术","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Proteomic insights into plant-endophyte interactions: advancing understanding of mutualistic symbiosis and plant resilience.","authors":"Shivam Chauhan, Poonam Kumari, Nikky Deepa, Jiya Chanotiya, Prabodh Kumar Trivedi, Akanksha Singh","doi":"10.1080/07388551.2026.2618190","DOIUrl":"10.1080/07388551.2026.2618190","url":null,"abstract":"<p><p>Endophytic microorganisms are a vital part of the plant microbiome, contributing significantly to the plant's growth, development, and stress tolerance. Proteomics investigations have significantly enhanced our comprehension of the interactions between plants and endophytes, illuminating the complex molecular mechanisms that govern these mutually beneficial relationships. The review aims to integrate the latest developments in proteomic research concerning endophyte-plant interactions, emphasizing on elucidating the molecular mechanisms that underlie the benefit imparted to the host plant by the symbionts. The special focus of the review is to discuss the proteome level changes happening at the early recognition events, primary and secondary metabolism, signaling pathways, and defense mechanisms. By underscoring critical proteomic signatures, the review aspires to offer insights into how these interactions enhance plant health, increase stress resilience, and promote overall growth. The article discusses the potential applications of proteomics in agriculture and environmental sciences, emphasizing its role in crop resilience against biotic and abiotic stresses, optimizing biocontrol strategies, and improving nutrient use efficiency. The article also highlights that despite the advancements, critical gaps persist including the necessity for a deeper understanding of the temporal dynamics of proteomic responses, the specificity of protein-protein interactions, and the influence of environmental factors on the proteome induced by the endophytes. The review concludes by proposing future directions for proteomics research in plant-endophyte interactions for developing a more comprehensive understanding of the intricate molecular dialogues for developing a more sustainable and resilient agricultural systems.</p>","PeriodicalId":10752,"journal":{"name":"Critical Reviews in Biotechnology","volume":" ","pages":"440-459"},"PeriodicalIF":8.7,"publicationDate":"2026-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147497778","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"工程技术","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Engineering non-ribosomal peptide synthesis: tuning the antibiotics engine of the microbial world.","authors":"Lucy Butler, Ali Raza Awan, Tom Ellis, Muhammad Safwan Akram","doi":"10.1080/07388551.2026.2615819","DOIUrl":"10.1080/07388551.2026.2615819","url":null,"abstract":"<p><p>Non-Ribosomal Peptide Synthetases produce chemically diverse peptides in nature, many of which have antimicrobial properties, providing an opportunity to use synthetic biology to fine tune them for pharmaceutical applications. Major challenges remain with total and semi-synthesis of these complex peptides with specific bioengineering methodologies being developed to increase low yields and enhance bioactivity. Here we review major advances in engineering non-ribosomal peptides with a focus on improvements made to achieve better yield and bioactivity. This can be achieved through: engineering precursor metabolites, altering metabolic flux, introducing strong promoters and regulators, and redirecting metabolism to biosynthetic gene clusters which can then be expressed natively or heterologously. We also review glycopeptide antibiotics as a promising opportunity for engineering through synthetic biology for the biosynthesis of novel non-ribosomal peptides.</p>","PeriodicalId":10752,"journal":{"name":"Critical Reviews in Biotechnology","volume":" ","pages":"501-521"},"PeriodicalIF":8.7,"publicationDate":"2026-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147343918","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"工程技术","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Spotlight on pectinase: a comprehensive review of large-scale production strategies.","authors":"Haruna Yahaya Abdullahi, Mohit Kumar, Santosh Kumar Mishra, Kavya Dashora, Soumya Pandit, Sonu Saini, Manikant Tripathi, Pallavi Mishra, Anoop Singh, Veeranna Channashettar, Gaurav Rajauria, Ramesh Chander Kuhad","doi":"10.1080/07388551.2025.2587149","DOIUrl":"10.1080/07388551.2025.2587149","url":null,"abstract":"<p><p>In recent years, pectinase, a vital enzyme in diverse manufacturing sectors, including: food and beverage industries, bioenergy, textile and paper industries, etc., has inspired the scientific community to delve its: sustainable, eco-friendly, efficient, and sufficient production. Pectinase demand is perpetually rising, requiring effective mass production solutions. This study examines the various sources and improvements made in the recent years at large-scale pectinase production. The article highlighted various fermentation strategies, agro-wastes, and types of bioreactor technology utilized for pectinase production. Further: statistical tools, research designs and optimization approaches, immobilization techniques, and purification and molecular engineering approaches were also explored, accounting pectinase production. The current work aims to provide the valuable insights for: researchers, academicians, industry stakeholders, and regulatory bodies, in advancing sustainable and efficient large-scale production of pectinase, thus, broadening and boosting pectinase production for the targeted applications.</p>","PeriodicalId":10752,"journal":{"name":"Critical Reviews in Biotechnology","volume":" ","pages":"297-317"},"PeriodicalIF":7.7,"publicationDate":"2026-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145899386","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"工程技术","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Development of microbial chassis for production of fungal natural products.","authors":"Junyang Wang, Xu Ji, Zhenlin Xin, Jinmiao Hu, Zihe Liu, Shuobo Shi","doi":"10.1080/07388551.2025.2608895","DOIUrl":"10.1080/07388551.2025.2608895","url":null,"abstract":"<p><p>Fungal natural products (FNPs) are an important class of natural medicines. Genome sequencing has uncovered an unexpectedly large number of silent biosynthetic gene clusters (BGCs) in fungi that hold potential for FNP production. However, activating silent BGCs in native hosts is hindered by undesirable traits, such as difficulty in genetic manipulation or exhibiting extremely low metabolite titers. With the development of synthetic biology, heterologous expression systems are increasingly becoming the preferred option to overcome these limitations. In this review, we first summarize the major structural classes of FNPs and the corresponding backbone enzymes. We then evaluate the key features of various microbial chassis and strategies employed for pathway refactoring to achieve efficient heterologous expression. Furthermore, we discuss optimization strategies that enhance pathway flux toward the target product and minimize by-product formation. These methodologies are essential for advancing heterologous platforms for FNP discovery and biosynthesis. Additionally, we analyze current challenges and propose solutions to further improve microbial chassis for more effective FNP production.</p>","PeriodicalId":10752,"journal":{"name":"Critical Reviews in Biotechnology","volume":" ","pages":"209-224"},"PeriodicalIF":8.7,"publicationDate":"2026-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146060664","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"工程技术","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Microalgae-based strategies for cadmium remediation: insights, challenges, and future directions.","authors":"Sawera Akram, Ge Guan, Beibei Hu, Muhammad Abu Bakar Saddique, Yitao Xi, Xiumei Luo, Maozhi Ren","doi":"10.1080/07388551.2025.2585681","DOIUrl":"10.1080/07388551.2025.2585681","url":null,"abstract":"<p><p>Cadmium (Cd²⁺) pollution possesses severe risks to human health and the ecosystem due to its high toxicity, persistence, and bioaccumulation potential. Conventional remediation methods, such as chemical precipitation, membrane filtration and ion exchange, are often costly, inefficient and unsustainable. In contrast, microalgae-based bioremediation has emerged as a promising approach due to its ability of biosorption, bioaccumulation and biotransformation. Microalgae possess unique metabolic and structural attributes, including: abundant extracellular metal binding sites, polymeric substances, intracellular chelators and the ability of Cd-nanoparticles (CdSeNPs, CdSNPs) formation enabling efficient Cd²⁺ sequestration and detoxification. Despite these advantages, large-scale application remains limited due to gaps in understanding of key regulatory mechanisms. This review highlights the detailed mechanism of the microalgae-based Cd²⁺ remediation process, identifies critical factors influencing remediation efficiency and potential microalgae strain's efficiency in Cd²⁺ removal. Furthermore, the utilization of genetic engineering for enhancing remediation efficiency by targeting key metal transporters, chelators, and stress-response pathways and potential candidate gene are also highlighted. These biotechnological advances and the understanding of the microalgae mediated remediation process presents a promise for a large scale efficient, sustainable Cd²⁺ bioremediation approach.</p>","PeriodicalId":10752,"journal":{"name":"Critical Reviews in Biotechnology","volume":" ","pages":"318-337"},"PeriodicalIF":7.7,"publicationDate":"2026-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145556467","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"工程技术","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Fungi in global hydrocarbon bioremediation: biodiversity, enzymatic pathways and technological innovations.","authors":"Cezar Júnio Coêlho Paranhos, Maria Júlia Lima Gonçalves, Edivaldo Ximenes Ferreira Filho, Luis Henrique Ferreira do Vale","doi":"10.1080/07388551.2025.2612089","DOIUrl":"10.1080/07388551.2025.2612089","url":null,"abstract":"<p><p>Fungi have emerged as powerful biological agents in the bioremediation of hydrocarbon-contaminated environments due to their robust enzymatic systems, adaptability, and ecological relevance. This review critically examines their potential, highlighting enzymatic mechanisms and technological innovations that underpin this sustainable approach. Case studies with: <i>Aspergillus sp.</i>, <i>Fusarium sp.</i>, <i>Paecilomyces sp.</i>, <i>Penicillium sp.</i>, and <i>Trametes sp.</i> demonstrate, through complex enzyme systems (laccases, lignin, manganese, and versatile peroxidases), the ability to degrade toxic compounds such as polycyclic aromatic hydrocarbons (PAHs) and BTEX, converting them into less harmful metabolites or even reusable by-products. Integrated strategies, including: biostimulation, bioaugmentation, microbial consortia, and the application of biosurfactants and bioemulsifiers, further enhance fungal efficiency in heterogeneous environments. Emerging innovations such as enzyme immobilization, myco-nanoremediation, and genetic engineering are discussed as promising solutions to overcome the inherent limitations of contaminant degradation under adverse conditions. Nevertheless, significant challenges remain, including the complexity of fungal metabolic pathways, gaps in proteomic regulation, and persistent obstacles in scaling and field reproducibility, which currently restrict large-scale commercial application. The alignment of fungal bioremediation with circular economy principles is emphasized, particularly the transformation of oil-derived pollutants into economically valuable bioproducts. Although fungal-based technologies remain underexplored commercially, especially in relation to regulatory frameworks and strategic partnerships, with this gap being especially evident in the Brazilian context, this review provides a robust foundation for advancing the application of fungi in sustainable environmental recovery. By integrating mechanistic insights with technological innovations and regulatory perspectives, it addresses a critical gap in the literature and outlines future directions for the field.</p>","PeriodicalId":10752,"journal":{"name":"Critical Reviews in Biotechnology","volume":" ","pages":"183-208"},"PeriodicalIF":8.7,"publicationDate":"2026-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146060733","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"工程技术","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}