Enzyme and Microbial Technology最新文献_第10页

Epoxidation of perillyl alcohol by engineered bacterial cytochrome P450 BM3 工程细菌细胞色素 P450 BM3 对过氧乙醇的环氧化作用。

IF 3.4 3区生物学

Enzyme and Microbial Technology Pub Date : 2024-07-29 DOI: 10.1016/j.enzmictec.2024.110487

Chan Mi Park , Gun Su Cha , Hae Chan Jeong , Yu-jin Lee , Jeong-Hoon Kim , Moon-Soo Chung , Sungbeom Lee , Chul-Ho Yun

{"title":"Epoxidation of perillyl alcohol by engineered bacterial cytochrome P450 BM3","authors":"Chan Mi Park , Gun Su Cha , Hae Chan Jeong , Yu-jin Lee , Jeong-Hoon Kim , Moon-Soo Chung , Sungbeom Lee , 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><sub>cat</sub> and <em>K</em><sub>m</sub> of M601 were 540 min<sup>−1</sup> 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}

引用次数: 0

Comparative biochemistry of PET hydrolase-carbohydrate-binding module fusion enzymes on a variety of PET substrates PET水解酶-碳水化合物结合模块融合酶在多种PET底物上的生物化学比较。

IF 3.4 3区生物学

Enzyme and Microbial Technology Pub Date : 2024-07-23 DOI: 10.1016/j.enzmictec.2024.110479

Andrew Philip Rennison , Andreas Prestel , Peter Westh , Marie Sofie Møller

{"title":"Comparative biochemistry of PET hydrolase-carbohydrate-binding module fusion enzymes on a variety of PET substrates","authors":"Andrew Philip Rennison , Andreas Prestel , Peter Westh , 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<sup>ICCG</sup> 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<sup>ICCG</sup> 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}

引用次数: 0

Identification and characterization of a novel thermostable PL7 alginate lyase from a submarine volcanic metagenomic library 从海底火山元基因组文库中鉴定新型恒温 PL7 藻酸盐裂解酶并确定其特征

IF 3.4 3区生物学

Enzyme and Microbial Technology Pub Date : 2024-07-21 DOI: 10.1016/j.enzmictec.2024.110486

Vasileios Tsopanakis , Elena Anastasiadou , Maria D. Mikkelsen , Anne S. Meyer , Ioannis V. Pavlidis

{"title":"Identification and characterization of a novel thermostable PL7 alginate lyase from a submarine volcanic metagenomic library","authors":"Vasileios Tsopanakis , Elena Anastasiadou , Maria D. Mikkelsen , Anne S. Meyer , Ioannis V. Pavlidis","doi":"10.1016/j.enzmictec.2024.110486","DOIUrl":"10.1016/j.enzmictec.2024.110486","url":null,"abstract":"<div><p>Seaweed biomass is as an abundant and renewable source of complex polysaccharides, including alginate which has a variety of applications. A sustainable method for exploiting alginate towards the production of valuable oligosaccharides is through enzymatic processing, using alginate lyases. Industrial refinement methods demand robust enzymes. Metagenomic libraries from extreme environments are a new source of unique enzymes with great industrial potential. Herein we report the identification of a new thermostable alginate lyase with only 58 % identity to known sequences, identified by mining a metagenomic library obtained from the hydrothermal vents of the volcano Kolumbo in the Aegean Sea (Kolumbo Alginate Lyase, KAlLy). Sequence analysis and biochemical characterization of KAlLy showed that this new alginate lyase is a Polysaccharide Lyase of family 7 (PL7) enzyme with endo- and exo-action on alginate and poly-mannuronic acid, with high activity at 60°C (56 ± 8 U/mg) and high thermostability (half-life time of 30 h at 50°C). The response surface methodology analysis revealed that the reaction optimum conditions with poly-mannuronic acid as substrate are 44°C, pH of 5.5 with 440 mM NaCl. This novel alginate lyase is a valuable addition to the toolbox of alginate modifying enzymes, due to its diverse sequence and its good thermal stability.</p></div>","PeriodicalId":11770,"journal":{"name":"Enzyme and Microbial Technology","volume":"180 ","pages":"Article 110486"},"PeriodicalIF":3.4,"publicationDate":"2024-07-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141736457","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}

引用次数: 0

High-efficient preparation of β-nicotinamide mononucleotides by crude enzymes cascade catalytic reaction 利用粗酶级联催化反应高效制备 β-烟酰胺单核苷酸。

IF 3.4 3区生物学

Enzyme and Microbial Technology Pub Date : 2024-07-20 DOI: 10.1016/j.enzmictec.2024.110482

Jiehu Liu , Runtian Huo , Huixian Fu , Shiheng Chen , Xueyi Qiao , Bo Xu , Zhaoyuan Zhang , Jing Wu , Lingqia Su

{"title":"High-efficient preparation of β-nicotinamide mononucleotides by crude enzymes cascade catalytic reaction","authors":"Jiehu Liu , Runtian Huo , Huixian Fu , Shiheng Chen , Xueyi Qiao , Bo Xu , Zhaoyuan Zhang , Jing Wu , Lingqia Su","doi":"10.1016/j.enzmictec.2024.110482","DOIUrl":"10.1016/j.enzmictec.2024.110482","url":null,"abstract":"<div><p>β-nicotinamide mononucleotide (β-NMN) is a key precursor of nicotinamide adenine dinucleotide, and becomes attractive in the nutrition and health care fields, but its enzymatic synthesis is expensive. In this study, a six-enzyme cascade catalytic system was constructed to produce β-NMN. Using D-ribose and nicotinamide as substrates, the β-NMN yield reached 97.5 % catalyzed by purified enzymes. Then, after knocking out the genes encoding proteins that consume β-NMN in <em>E. coli</em> BL21(DE3), the similar β-NMN yield, 97.2 %, using the crude enzymes could be also obtained. After that, β-NMN synthesis was performed under increased substrate concentration, and 'modular' crude enzymes cascade catalytic reaction system was proposed to reduce the inhibition of polyphosphate on ribose-phosphate diphosphokinase activity, and the β-NMN yield reached 78.4 % at 10 mM D-ribose, which is 1.82 times of that in 'one-pot' reaction and represents the highest β-NMN preparation level with phosphoribosylpyrophosphate as the core reported till now.</p></div>","PeriodicalId":11770,"journal":{"name":"Enzyme and Microbial Technology","volume":"180 ","pages":"Article 110482"},"PeriodicalIF":3.4,"publicationDate":"2024-07-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141765748","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}

引用次数: 0

Efficiency enhancement in Aspergillus niger α-L-rhamnosidase reverse hydrolysis by using a tunnel site rational design strategy 利用隧道位点合理设计策略提高黑曲霉α-L-鼠李糖酶反向水解的效率

IF 3.4 3区生物学

Enzyme and Microbial Technology Pub Date : 2024-07-19 DOI: 10.1016/j.enzmictec.2024.110484

Yanling Lin , Yuchen Cai , Han Li , Lijun Li , Zedong Jiang , Hui Ni

{"title":"Efficiency enhancement in Aspergillus niger α-L-rhamnosidase reverse hydrolysis by using a tunnel site rational design strategy","authors":"Yanling Lin , Yuchen Cai , Han Li , Lijun Li , Zedong Jiang , Hui Ni","doi":"10.1016/j.enzmictec.2024.110484","DOIUrl":"10.1016/j.enzmictec.2024.110484","url":null,"abstract":"<div><p>There has been ongoing interest in improving the efficiency of glycoside hydrolase for synthesizing glycoside compounds through protein engineering, given the potential applications of glycoside compounds. In this study, a strategy of modifying the substrate access tunnel was proposed to enhance the efficiency of reverse hydrolysis catalyzed by <em>Aspergillus niger</em> α-L-rhamnosidase. Analysis of the tunnel dynamics identified Tyr299 as a key modifiable residue in the substrate access tunnel. The location of Tyr299 was near the enzyme surface and at the outermost end of the substrate access tunnel, suggested its role in substrate recognition and throughput. Based on the properties of side chains, six mutants were designed and expressed by <em>Pichia pastoris</em>. Compared to WT, the reverse hydrolysis efficiencies of mutants Y299P and Y299W were increased by 21.3 % and 11.1 %, respectively. The calculation results of binding free energy showed that the binding free energy was inversely proportional to the reverse hydrolysis efficiency. Further, when binding free energy levels were comparable, the mutants with shorter side chains displayed a higher reverse hydrolysis efficiency. These results proved that substrate access tunnel modification was an effective method to improve the reverse hydrolysis efficacy of α-L-rhamnosidase and also provided new insights for modifying other glycoside hydrolases.</p></div>","PeriodicalId":11770,"journal":{"name":"Enzyme and Microbial Technology","volume":"180 ","pages":"Article 110484"},"PeriodicalIF":3.4,"publicationDate":"2024-07-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141852485","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}

引用次数: 0

Biocatalytic approach for the synthesis of chiral alcohols for the development of pharmaceutical intermediates and other industrial applications: A review 生物催化方法合成手性醇，用于开发医药中间体和其他工业应用：综述

IF 3.4 3区生物学

Enzyme and Microbial Technology Pub Date : 2024-07-17 DOI: 10.1016/j.enzmictec.2024.110483

Mohd Naim , Mohd Fazli Mohammat , Putri Nur Arina Mohd Ariff , Mohamad Hekarl Uzir

{"title":"Biocatalytic approach for the synthesis of chiral alcohols for the development of pharmaceutical intermediates and other industrial applications: A review","authors":"Mohd Naim , Mohd Fazli Mohammat , Putri Nur Arina Mohd Ariff , Mohamad Hekarl Uzir","doi":"10.1016/j.enzmictec.2024.110483","DOIUrl":"10.1016/j.enzmictec.2024.110483","url":null,"abstract":"<div><p>Biocatalysis has emerged as a strong tool for the synthesis of active pharmaceutical ingredients (APIs). In the early twentieth century, whole cell biocatalysis was used to develop the first industrial biocatalytic processes, and the precise work of enzymes was unknown. Biocatalysis has evolved over the years into an essential tool for modern, cost-effective, and sustainable pharmaceutical manufacturing. Meanwhile, advances in directed evolution enable the rapid production of process-stable enzymes with broad substrate scope and high selectivity. Large-scale synthetic pathways incorporating biocatalytic critical steps towards >130 APIs of authorized pharmaceuticals and drug prospects are compared in terms of steps, reaction conditions, and scale with the corresponding chemical procedures. This review is designed on the functional group developed during the reaction forming alcohol functional groups. Some important biocatalyst sources, techniques, and challenges are described. A few APIs and their utilization in pharmaceutical drugs are explained here in this review. Biocatalysis has provided shorter, more efficient, and more sustainable alternative pathways toward existing small molecule APIs. Furthermore, non-pharmaceutical applications of biocatalysts are also mentioned and discussed. Finally, this review includes the future outlook and challenges of biocatalysis. In conclusion, Further research and development of promising enzymes are required before they can be used in industry.</p></div>","PeriodicalId":11770,"journal":{"name":"Enzyme and Microbial Technology","volume":"180 ","pages":"Article 110483"},"PeriodicalIF":3.4,"publicationDate":"2024-07-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141732151","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}

引用次数: 0

Enhanced L-theanine production through semi-rational design of γ-glutamylmethylamide synthetase from Methylovorus mays 通过对 Methylovorus mays 的 γ-谷氨酰甲酰胺合成酶进行半合理设计，提高 L-茶氨酸的产量。

IF 3.4 3区生物学

Enzyme and Microbial Technology Pub Date : 2024-07-17 DOI: 10.1016/j.enzmictec.2024.110481

Chao Fan , Jiakun Qi , Yunhan Cong , Chunzhi Zhang

引用次数: 0

Osmoregulation by choline-based deep eutectic solvent induces electroactivity in Bacillus subtilis biofilms 胆碱深共晶溶剂的渗透调节作用诱导枯草芽孢杆菌生物膜的电活性

IF 3.4 3区生物学

Enzyme and Microbial Technology Pub Date : 2024-07-16 DOI: 10.1016/j.enzmictec.2024.110485

Neda Eghtesadi , Kayode Olaifa , Tri T. Pham , Vito Capriati , Obinna M. Ajunwa , Enrico Marsili

{"title":"Osmoregulation by choline-based deep eutectic solvent induces electroactivity in Bacillus subtilis biofilms","authors":"Neda Eghtesadi , Kayode Olaifa , Tri T. Pham , Vito Capriati , Obinna M. Ajunwa , Enrico Marsili","doi":"10.1016/j.enzmictec.2024.110485","DOIUrl":"10.1016/j.enzmictec.2024.110485","url":null,"abstract":"<div><p>Gram-positive <em>Bacillus subtilis</em> is a model organism for the biotechnology industry and has recently been characterized as weakly electroactive in both planktonic cultures and biofilms. Increasing the extracellular electron transfer (EET) rate in <em>B. subtilis</em> biofilms will help to develop an efficient microbial electrochemical technology (MET) and improve the bioproduction of high-value metabolites under electrofermentative conditions. In our previous work, we have shown that the addition of compatible solute precursors such as choline chloride (ChCl) to the growth medium formulation increases current output and biofilm formation in <em>B. subtilis</em>. In this work, we utilized a low-carbon tryptone yeast extract medium with added salts to further expose <em>B. subtilis</em> to salt stress and observe the osmoregulatory and/or nutritional effects of a D-sorbitol/choline chloride (ChCl) (1:1 mol mol<sup>−1</sup>) deep eutectic solvents (DESs) on the electroactivity of the formed biofilm. The results show that ChCl and D-sorbitol alleviate the osmotic stress induced by the addition of NaH<sub>2</sub>PO<sub>4</sub> and KH<sub>2</sub>PO<sub>4</sub> salts and boost biofilm production. This is probably due to the osmoprotective effect of ChCl, a precursor of the osmoprotectant glycine betaine, and the induction of electroactive exopolymeric substances within the <em>B. subtilis</em> biofilm. Since high ionic strength media are commonly used in microbial biotechnology, the combination of ChCl-containing DESs and salt stress could enhance biofilm-based electrofermentation processes that bring significant benefits for biotechnological applications.</p></div>","PeriodicalId":11770,"journal":{"name":"Enzyme and Microbial Technology","volume":"180 ","pages":"Article 110485"},"PeriodicalIF":3.4,"publicationDate":"2024-07-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.sciencedirect.com/science/article/pii/S0141022924000929/pdfft?md5=407aeace407d0e7ef745e4781151fc3c&pid=1-s2.0-S0141022924000929-main.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141709890","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}

引用次数: 0

Rational design of short-chain dehydrogenase DHDR for efficient synthesis of (S)-equol 合理设计短链脱氢酶 DHDR 以高效合成 (S)-equol

IF 3.4 3区生物学

Enzyme and Microbial Technology Pub Date : 2024-07-16 DOI: 10.1016/j.enzmictec.2024.110480

Weichuang Qin , Lujia Zhang , Yichen Yang , Wei Zhou , Shuting Hou , Jie Huang , Bei Gao

{"title":"Rational design of short-chain dehydrogenase DHDR for efficient synthesis of (S)-equol","authors":"Weichuang Qin , Lujia Zhang , Yichen Yang , Wei Zhou , Shuting Hou , Jie Huang , Bei Gao","doi":"10.1016/j.enzmictec.2024.110480","DOIUrl":"10.1016/j.enzmictec.2024.110480","url":null,"abstract":"<div><p>(<em>S</em>)-equol, the most influential metabolite of daidzein in <em>vivo</em>, has aroused great attention due to the excellent biological activities. Although existing studies have accomplished the construction of its heterologous synthetic pathway in the context of anaerobicity and inefficiency of natural strains, the low productivity of (<em>S</em>)-equol limits its industrial application. Here, rational design strategies based on decreasing the pocket steric hindrance and fine-tuning the pocket microenvironment to systematically redesign the binding pocket of enzyme were developed and processed to the rate-limiting enzyme dihydrodaidzein reductase in (<em>S</em>)-equol synthesis. After iterative combinatorial mutagenesis, an effective mutant S118G/T169A capable of significantly increasing (<em>S</em>)-equol yield was obtained. Computational analyses illustrated that the main reason of the increased activity relied on the decreased critical distance and more stable interacting conformation. Then, the reaction optimization was performed, and the recombinant <em>Escherichia coli</em> whole-cell biocatalyst harboring S118G/T169A enabled the efficient conversion of 2 mM daidzein to (<em>S</em>)-equol, achieving conversion rate of 84.5 %, which was 2.9 times higher than that of the parental strain expressing wide type dihydrodaidzein reductase. This study provides an effective idea and a feasible method for enzyme modification and whole-cell catalytic synthesis of (<em>S</em>)-equol, and will greatly accelerate the process of industrial production.</p></div>","PeriodicalId":11770,"journal":{"name":"Enzyme and Microbial Technology","volume":"180 ","pages":"Article 110480"},"PeriodicalIF":3.4,"publicationDate":"2024-07-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141697498","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}

引用次数: 0

Debridement efficacy of serine protease and formulated cream by In Vitro assessment against artificial wound eschar 通过体外评估丝氨酸蛋白酶和配制膏对人工伤口焦痂的清创功效

IF 3.4 3区生物学

Enzyme and Microbial Technology Pub Date : 2024-07-11 DOI: 10.1016/j.enzmictec.2024.110478

Julia Yunus, Haryati Jamaluddin, Wan Rosmiza Zana Wan Dagang

{"title":"Debridement efficacy of serine protease and formulated cream by In Vitro assessment against artificial wound eschar","authors":"Julia Yunus, Haryati Jamaluddin, Wan Rosmiza Zana Wan Dagang","doi":"10.1016/j.enzmictec.2024.110478","DOIUrl":"10.1016/j.enzmictec.2024.110478","url":null,"abstract":"<div><p>Chronic wounds typically comprise of necrotic tissue and dried secretions, often culminating in the formation of a thick and tough layer of dead skin known as eschar. Removal of eschar is imperative to facilitate wound healing. Conventional approach for eschar removal involves surgical excision and grafting, which can be traumatic and frequently leads to viable tissue damage. There has been growing interest in the use of enzymatic agents for a gentler approach to debridement, utilizing proteolytic enzymes. In this study, a purified intracellular recombinant serine protease from <em>Bacillus</em> sp. (SPB) and its cream formulation were employed to evaluate their ability to degrade artificial wound eschar; composed of collagen, fibrin, and elastin. Degradation was assessed based on percentage weight reduction of eschar biomass, analysis via sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE), and scanning electron microscopy (SEM). Both SPB and its cream formulation were able to degrade up to 50 % artificial wound eschar, with the SPB cream maintaining its degradation efficiency for up to 24 hours. Additionally, the SPB-based cream demonstrated the ability to hydrolyze proteinaceous components of eschars individually (fibrin and collagen) as determined through qualitative assessment. These findings suggest that SPB holds promise for the debridement of wound eschar.</p></div>","PeriodicalId":11770,"journal":{"name":"Enzyme and Microbial Technology","volume":"180 ","pages":"Article 110478"},"PeriodicalIF":3.4,"publicationDate":"2024-07-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141695010","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}

引用次数: 0