Enzyme and Microbial Technology最新文献

{"title":"A directional electrode separator improves anodic biofilm current density in a well-mixed single-chamber bioelectrochemical system","authors":"Md Monzurul Islam Anoy ,&nbsp;Eric Allen Hill ,&nbsp;Marci Ranae Garcia ,&nbsp;Won-Jun Kim ,&nbsp;Alexander S. Beliaev ,&nbsp;Haluk Beyenal","doi":"10.1016/j.enzmictec.2024.110502","DOIUrl":"10.1016/j.enzmictec.2024.110502","url":null,"abstract":"<div><p>In this study, a directional electrode separator (DES) was designed and incorporated into a single-chamber bioelectrochemical system (BES) to reduce migration and reoxidation of hydrogen. This issue arises when H₂, generated at the cathode, travels to the anode where anodic biofilms use H₂. To test the feasibility of our design, a 3D-printed BES reactor equipped with a DES was inoculated with anaerobic digestor granules and operated under fed-batch conditions using fermented corn stover effluent. The DES equipped reactor achieved significantly higher current densities (∼53 A/m²) compared to a conventional single-chamber BES without a separator (∼16 A/m²), showing a 3.3 times improvement. Control abiotic electrochemical experiments revealed that the DES exhibited significantly higher proton conductivity (456±127 µS/mm) compared to a proton exchange membrane (67±21 µS/mm) with a statistical significance of P=0.03. The DES also effectively reduced H₂ migration to the anode by 21-fold relative to the control. Overall, incorporating a DES in a single-chamber BES enhanced anodic current density by reducing H₂ migration to the anode.</p></div>","PeriodicalId":11770,"journal":{"name":"Enzyme and Microbial Technology","volume":"180 ","pages":"Article 110502"},"PeriodicalIF":3.4,"publicationDate":"2024-08-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142098426","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Characterization of a secondary palmitoleoyltransferase of lipid A in Vibrio parahaemolyticus","authors":"Danyang Huang ,&nbsp;Lingyan Chen ,&nbsp;Zhe Wang ,&nbsp;Fenfang He ,&nbsp;Xinrui Zhang ,&nbsp;Xiaoyuan Wang","doi":"10.1016/j.enzmictec.2024.110504","DOIUrl":"10.1016/j.enzmictec.2024.110504","url":null,"abstract":"<div><p>The detection of pathogenicity and immunogenicity in <em>Vibrio parahaemolyticus</em> poses a significant challenge due to its threat to human health and food safety, which is strongly correlated with lipid A. Lipid A, a critical component found in most Gram-negative bacteria, functions as a hydrophobic anchor for lipopolysaccharide. <em>V. parahaemolyticus</em> synthesizes multiple lipid A species with various secondary acyl chains. In this study, a secondary acyltransferase of lipid A encoded by <em>VP_RS08405</em> in <em>V. parahaemolyticus</em> was identified. Based on sequence alignment analysis, <em>V. parahaemolyticus VP_RS08405</em> has high homology to <em>E. coli lpxL</em>, <em>lpxM</em> and <em>lpxP</em> which encode the three secondary acyltransferases of lipid A. Therefore, <em>V. parahaemolyticus VP_RS08405</em> was cloned into pBAD33, and the resulting pB08405 was introduced in <em>E. coli</em> mutants WHL00 in which <em>lpxL</em> was deleted, WHM00 in which <em>lpxM</em> was deleted, WHP00 in which <em>lpxP</em> was deleted, and WH300 in which <em>lpxL</em>, <em>lpxM</em> and <em>lpxP</em> were deleted. The recombinant strains WHL00/pB08405, WHM00/pB08405, WHP00/pB08405, WH300/pB08405, as well as their vector controls, were grown at normal and low temperatures. Lipid A species were isolated from the above strains and analyzed by using high-performance liquid chromatography-tandem mass spectrometry and thin-layer chromatography. After comparing the secondary acyl alterations of lipid A from different recombinant strains, it is concluded that VP_RS08405 specifically catalyzed the addition of a palmitoleate to the 2′-position of lipid A and its activity is not temperature-sensitive. In addition, to determine the dependence of VP_RS08405 on Kdo, <em>VP_RS08405</em> was overexpressed in <em>E. coli</em> mutants WH001 in which <em>waaA</em> was deleted, and WH400 in which <em>waaA</em>, <em>lpxL</em>, <em>lpxM</em> and <em>lpxP</em> were deleted. Lipid A species were isolated from WH001/pB08405 and WH400/pB08405, and analyzed. The results show that the function of VP_RS08405 is Kdo-dependent. These findings provide a better understanding of the structural diversity of lipid A in <em>V. parahaemolyticus</em>.</p></div>","PeriodicalId":11770,"journal":{"name":"Enzyme and Microbial Technology","volume":"180 ","pages":"Article 110504"},"PeriodicalIF":3.4,"publicationDate":"2024-08-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142077114","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Enhanced glucose-1-phosphate production from corn stover using cellulases with reduced β-glucosidase activity via Trbgl1 gene knockout in Trichoderma reesei Rut C30","authors":"Xiaoqin Ran,&nbsp;Yushan Gao,&nbsp;Xiao He,&nbsp;Zancheng Wang,&nbsp;Yi Mo,&nbsp;Yonghao Li","doi":"10.1016/j.enzmictec.2024.110503","DOIUrl":"10.1016/j.enzmictec.2024.110503","url":null,"abstract":"<div><p>The scarcity of cellulases with low β-glucosidase activity poses a significant technological challenge in precisely controlling the partial hydrolysis of lignocellulose to cellobiose, crucial for producing high-value chemicals such as starch, inositol, and NMN. <em>Trichoderma reesei</em> is a primary strain in cellulase production. Therefore, this study targeted the critical β-glucosidase gene, <em>Trbgl1</em>, resulting in over an 86 % reduction in β-glucosidase activity. However, cellulase production decreased by 19.2 % and 20.3 % with lactose or cellulose inducers, respectively. Notably, transcript levels of cellulase genes and overall yield remained unaffected with an inducer containing sophorose. This indicates that β-glucosidase BGL1 converts lactose or cellulose to sophorose through transglycosylation activity, inducing cellulase gene transcription. The resulting enzyme cocktail, comprising recombinant cellulase and cellobiose phosphorylase, was applied for corn stover hydrolysis, resulting in a 24.3 % increase in glucose-1-phosphate yield. These findings provide valuable insights into obtaining enzymes suitable for the high-value utilization of lignocellulose.</p></div>","PeriodicalId":11770,"journal":{"name":"Enzyme and Microbial Technology","volume":"180 ","pages":"Article 110503"},"PeriodicalIF":3.4,"publicationDate":"2024-08-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142087989","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Enhancing the thermal stability and activity of zearalenone lactone hydrolase to promote zearalenone degradation via semi-rational design","authors":"Xue Jiang ,&nbsp;Sana Tehreem ,&nbsp;Kashif Rahim ,&nbsp;Meixing Wang ,&nbsp;Pan Wu ,&nbsp;Guimin Zhang","doi":"10.1016/j.enzmictec.2024.110499","DOIUrl":"10.1016/j.enzmictec.2024.110499","url":null,"abstract":"<div><p>Zearalenone (ZEN) is a fungal toxin produced by <em>Fusarium exospore</em>, which poses a significant threat to both animal and human health due to its reproductive toxicity. Removing ZEN through ZEN lactonase is currently the most effective method reported, however, all published ZEN lactonases suffer from the poor thermal stability, losing almost all activity after 10 min of treatment at 55℃. In this study, we heterologously expressed ZHD11A from <em>Phialophora macrospora</em> and engineered it via semi-rational design. A mutant I160Y-G242S that can retain about 40 % residual activity at 55℃ for 10 min was obtained, which is the most heat-tolerant ZEN hydrolase reported to date. Moreover, the specific activity of the I160Y-G242S was also elevated 2-fold compared to ZHD11A from 220 U/mg to 450 U/mg, which is one of the most active ZEN lactonses reported. Dynamics analysis revealed that the decreased flexibility of the main-chain carbons contributes to increased thermal stability and the improved substrate binding affinity and catalytic turnover contribute to enhanced activity of variant I160Y-G242S. In all, the mutant I160Y-G242S is an excellent candidate for the industrial application of ZEN degradation.</p></div>","PeriodicalId":11770,"journal":{"name":"Enzyme and Microbial Technology","volume":"180 ","pages":"Article 110499"},"PeriodicalIF":3.4,"publicationDate":"2024-08-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142079728","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Unveiling the crystal structure of thermostable dienelactone hydrolase exhibiting activity on terephthalate esters","authors":"Dnane Vieira Almeida ,&nbsp;Iara Ciancaglini ,&nbsp;Ana Luiza Hernandes Sandano ,&nbsp;Ellen K.B. Roman ,&nbsp;Viviane Brito Andrade ,&nbsp;Ana Bárbara Nunes ,&nbsp;Robson Tramontina ,&nbsp;Viviam Moura da Silva ,&nbsp;Frank Gabel ,&nbsp;Thamy L.R. Corrêa ,&nbsp;André Damasio ,&nbsp;João Renato Carvalho Muniz ,&nbsp;Fabio Marcio Squina ,&nbsp;Wanius Garcia","doi":"10.1016/j.enzmictec.2024.110498","DOIUrl":"10.1016/j.enzmictec.2024.110498","url":null,"abstract":"<div><p>Dienelactone hydrolase (DLH) is one of numerous hydrolytic enzymes with an α/β-hydrolase fold, which catalyze the hydrolysis of dienelactone to maleylacetate. The DLHs share remarkably similar tertiary structures and a conserved arrangement of catalytic residues. This study presents the crystal structure and comprehensive functional characterization of a novel thermostable DLH from the bacterium <em>Hydrogenobacter thermophilus</em> (<em>Ht</em>DLH). The crystal structure of the <em>Ht</em>DLH, solved at a resolution of about 1.67 Å, exhibits a canonical α/β-hydrolase fold formed by eight β-sheet strands in the core, with one buried α-helix and six others exposed to the solvent. The structure also confirmed the conserved catalytic triad of DHLs formed by Cys121, Asp170, and His202 residues. The <em>Ht</em>DLH forms stable homodimers in solution. Functional studies showed that <em>Ht</em>DLH has the expected esterase activity over esters with short carbon chains, such as <em>p-</em>nitrophenyl acetate, reaching optimal activity at pH 7.5 and 70 °C. Furthermore, <em>Ht</em>DLH maintains more than 50 % of its activity even after incubation at 90 °C for 16 h. Interestingly, <em>Ht</em>DLH exhibits catalytic activity towards polyethylene terephthalate (PET) monomers, including bis-1,2-hydroxyethyl terephthalate (BHET) and 1-(2-hydroxyethyl) 4-methyl terephthalate, as well as other aliphatic and aromatic esters. These findings associated with the lack of activity on amorphous PET indicate that <em>Ht</em>DLH has characteristic of a BHET-degrading enzyme. This work expands our understanding of enzyme families involved in PET degradation, providing novel insights for plastic biorecycling through protein engineering, which could lead to eco-friendly solutions to reduce the accumulation of plastic in landfills and natural environments.</p></div>","PeriodicalId":11770,"journal":{"name":"Enzyme and Microbial Technology","volume":"180 ","pages":"Article 110498"},"PeriodicalIF":3.4,"publicationDate":"2024-08-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142047994","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Substrate specificity of commercial lipases activated by a hydration–aggregation pretreatment in anhydrous esterification reactions","authors":"Takashi Kuroiwa,&nbsp;Maho Katayama,&nbsp;Kazuki Uemoto,&nbsp;Akihiko Kanazawa","doi":"10.1016/j.enzmictec.2024.110497","DOIUrl":"10.1016/j.enzmictec.2024.110497","url":null,"abstract":"<div><p>Substrate specificity in non-aqueous esterification catalyzed by commercial lipases activated by hydration–aggregation pretreatment was investigated. Four microbial lipases from <em>Rhizopus japonicus</em>, <em>Burkholderia cepacia</em>, <em>Rhizomucor miehei</em>, and <em>Candida antarctica</em> (fraction B) were used to study the effect of the carbon chain length of saturated fatty acid substrates on the esterification activity with methanol in <em>n</em>-hexane. Hydration–aggregation pretreatment had an activation effect on all lipases used, and different chain length dependencies of esterification activity for lipases from different origins were demonstrated. The effects of various acidic substrates with different degrees of unsaturation, aromatic rings, and alcohol substrates with different carbon chain lengths on esterification activity were examined using <em>R. japonicus</em> lipase, which demonstrated the most remarkable activity enhancement after hydration–aggregation pretreatment. Furthermore, in the esterification of myristic acid with methanol catalyzed by the hydrated–aggregated <em>R. japonicus</em> lipase, maximum reaction rate (5.43 × 10⁻⁵ mmol/(mg-biocat min)) and Michaelis constants for each substrate (48.5 mM for myristic acid, 24.7 mM for methanol) were determined by kinetic analysis based on the two-substrate Michaelis-Menten model.</p></div>","PeriodicalId":11770,"journal":{"name":"Enzyme and Microbial Technology","volume":"180 ","pages":"Article 110497"},"PeriodicalIF":3.4,"publicationDate":"2024-08-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141998286","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Operation parameters and temperature affected sludge microbial metabolisms: An integrated perspective considering extracellular polymeric substances, soluble microbial products, biomass quantities, and community shifts","authors":"Xiaoliang Li ,&nbsp;Weiqi Luo ,&nbsp;Xinyu Yang ,&nbsp;Tong Liu ,&nbsp;Gang Tang ,&nbsp;Mengjing Guo ,&nbsp;Min Wang ,&nbsp;Xing Zheng","doi":"10.1016/j.enzmictec.2024.110496","DOIUrl":"10.1016/j.enzmictec.2024.110496","url":null,"abstract":"<div><p>Effluent soluble microbial products (SMP) and extracellular polymeric substances (EPS) are significant organics that pose challenges to advanced treatment processes. However, their production, transformation, and decomposition remain unclear due to their heterogeneity and the combined effects of environmental and operational factors. In this work, we investigated the impact of solids retention time (SRT), hydraulic retention time (HRT), and temperature on the changes in effluent SMP, with the consideration of the co-variation of EPS, sludge biomass, and community structures. Results show that longer SRT increased the biomass and relative abundance of functional microorganisms such as Myxococcota, Actinobacteria, and <em>Terrimonas</em>, which hindered EPS-to-SMP turnover and/or facilitated SMP consumption. This resulted in the accumulation of EPS and lower SMP concentrations at the beginning of the SRT adjustment. Both longer and shorter HRT (12 h and 8 h) led to increased SMP concentration, with the shorter HRT nearly doubling it (from approximately 6 to 12 mg/L), especially in terms of its protein and polysaccharide contents. Lower temperatures increased the SMP concentration and the relative abundance of Proteobacteria (including <em>Zoogloea</em>, the most dominant phylum and genus, relative abundance from 15.7 % to 61.1 %) while decreasing fluorescent EPS components, indicating the key role of Proteobacteria in SMP production and fluorescent EPS-to-SMP transformation. The results provided key insights into how changes in operational/environmental parameters impact sludge-EPS-SMP interactions, which could benefit the model development and operational optimization of activated sludge systems. This study also highlighted the important role of the sludge community in the EPS/SMP dynamics.</p></div>","PeriodicalId":11770,"journal":{"name":"Enzyme and Microbial Technology","volume":"180 ","pages":"Article 110496"},"PeriodicalIF":3.4,"publicationDate":"2024-08-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142044626","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Substrate specificity modification of paraben hydrolase and tannase from Aspergillus oryzae","authors":"Michiko Hakoda,&nbsp;Tomoe Kato,&nbsp;Chihiro Takahashi,&nbsp;Yoshihito Shiono,&nbsp;Takuya Koseki","doi":"10.1016/j.enzmictec.2024.110495","DOIUrl":"10.1016/j.enzmictec.2024.110495","url":null,"abstract":"<div><p>Paraben hydrolase and tannase catalyze the hydrolysis of parabens (4-hydroxybenzoic acid esters) and gallic acid (3,4,5-trihydroxybenzoic acid) esters, respectively. Paraben hydrolase (<em>Ao</em>PrbA) and tannase (<em>Ao</em>TanB) from <em>Aspergillus oryzae</em> belong to the tannase family in the ESTHER database. However, the substrate specificities of <em>Ao</em>PrbA and <em>Ao</em>TanB are narrow. Based on structural information of <em>Aspergillus niger</em> tannase (PDB code 7k4o), we constructed five single variants of <em>Ao</em>PrbA (Thr200Glu, Phe231Gln, Leu232Gln, Ile361Tyr, and Leu428Ser) and four of <em>Ao</em>TanB (Glu203Asp, Glu203Thr, His237Ala, and Ser440Leu) to investigate substrate discrimination between <em>Ao</em>PrbA and <em>Ao</em>TanB. Each variant was expressed in <em>Pichia pastoris</em> and were purified from the culture supernatant. Five purified variants of <em>Ao</em>PrbA and four variants of <em>Ao</em>TanB showed reduced paraben hydrolase and tannase activities compared with <em>Ao</em>PrbA and <em>Ao</em>TanB wild types, respectively. Interestingly, the <em>Ao</em>PrbA wild type did not hydrolyze gallic acid methyl ester, whereas the Thr200Glu, Leu232Gln, and Leu428Ser variants did, indicating that these three variants acquired tannase activity. In particular, the Leu428Ser variant exhibited considerably greater hydrolysis of gallic acid and protocatechuic acid methyl esters. Meanwhile, the <em>Ao</em>TanB wild type, and Glu203Asp, His237Ala and Ser440Leu variants hydrolyzed the protocatechuate methyl and 4-hydroxybenzoate ethyl esters; however, the Glu203Thr variant did not hydrolyze above-mentioned substrates. Additionally, the ratio of paraben hydrolase activity to tannase activity in Ser440Leu was markedly elevated.</p></div>","PeriodicalId":11770,"journal":{"name":"Enzyme and Microbial Technology","volume":"180 ","pages":"Article 110495"},"PeriodicalIF":3.4,"publicationDate":"2024-08-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141912266","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Epoxidation of perillyl alcohol by engineered bacterial cytochrome P450 BM3","authors":"Chan Mi Park ,&nbsp;Gun Su Cha ,&nbsp;Hae Chan Jeong ,&nbsp;Yu-jin Lee ,&nbsp;Jeong-Hoon Kim ,&nbsp;Moon-Soo Chung ,&nbsp;Sungbeom Lee ,&nbsp;Chul-Ho Yun","doi":"10.1016/j.enzmictec.2024.110487","DOIUrl":"10.1016/j.enzmictec.2024.110487","url":null,"abstract":"<div><p>Perillyl alcohol (POH) is a secondary metabolite of plants. POH and its derivatives are known to be effective as an anticancer treatment. In this study, oxidative derivatives of POH, which are difficult to synthesize chemically, were synthesized using the engineered bacterial cytochrome P450 BM3 (CYP102A1) as a biocatalyst. The activity of wild-type (WT) CYP102A1 and 29 engineered enzymes toward POH was screened using a high-performance liquid chromatography. They produced one major product. Among them, the engineered CYP102A1 M601 mutant with seven mutations (R47L/F81I/F87V/E143G/L150F/L188Q/E267V) showed the highest conversion, 6.4-fold higher than the WT. Structure modeling using AlphFold2 and PyMoL suggests that mutations near the water channel may be responsible for the increased catalytic activity of the M601 mutant. The major product was identified as a POH-8,9-epoxide by gas chromatography-mass spectrometry and nuclear magnetic resonance analysis. The optimal temperature and pH for the product formation were 35 °C and pH 7.4, respectively. The <em>k</em>_cat and <em>K</em>_m of M601 were 540 min⁻¹ and 2.77 mM, respectively. To improve POH-8,9-epoxide production, substrate concentration and reaction time were optimized. The optimal condition for POH-8,9-epoxide production by M601 was 5.0 mM POH, pH 7.4, 35 ℃, and 6 h reaction, which produced the highest concentration of 1.72 mM. Therefore, the biosynthesis of POH-8,9-epoxide using M601 as a biocatalyst is suggested to be an efficient and sustainable synthetic process that can be applied to chemical and pharmaceutical industries.</p></div>","PeriodicalId":11770,"journal":{"name":"Enzyme and Microbial Technology","volume":"180 ","pages":"Article 110487"},"PeriodicalIF":3.4,"publicationDate":"2024-07-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141855161","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Comparative biochemistry of PET hydrolase-carbohydrate-binding module fusion enzymes on a variety of PET substrates","authors":"Andrew Philip Rennison ,&nbsp;Andreas Prestel ,&nbsp;Peter Westh ,&nbsp;Marie Sofie Møller","doi":"10.1016/j.enzmictec.2024.110479","DOIUrl":"10.1016/j.enzmictec.2024.110479","url":null,"abstract":"<div><p>Enzyme-driven recycling of PET has now become a fully developed industrial process. With the right pre-treatment, PET can be completely depolymerized within workable timeframes. This has been realized due to extensive research conducted over the past decade, resulting in a large set of engineered PET hydrolases. Among various engineering strategies to enhance PET hydrolases, fusion with binding domains has been used to tune affinity and boost activity of the enzymes. While fusion enzymes have demonstrated higher activity in many cases, these results are primarily observed under conditions that would not be economically viable at scale. Furthermore, the wide variation in PET substrates, conditions, and combinations of PET hydrolases and binding domains complicates direct comparisons. Here, we present a self-consistent and thorough analysis of two leading PET hydrolases, LCC^ICCG and PHL7. Both enzymes were evaluated both without and with a substrate-binding domain across a range of industrially relevant PET substrates. We demonstrate that the presence of a substrate-binding module does not significantly affect the affinity of LCC^ICCG and PHL7 for PET. However, significant differences exist in how the fusion enzymes act on different PET substrates and solid substrate loading, ranging from a 3-fold increase in activity to a 6-fold decrease. These findings could inform the tailoring of enzyme choice to different industrial scenarios.</p></div>","PeriodicalId":11770,"journal":{"name":"Enzyme and Microbial Technology","volume":"180 ","pages":"Article 110479"},"PeriodicalIF":3.4,"publicationDate":"2024-07-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.sciencedirect.com/science/article/pii/S0141022924000863/pdfft?md5=03ce257e9b85718bb99039850b33e78d&pid=1-s2.0-S0141022924000863-main.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141757881","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}