Preparative Biochemistry & Biotechnology最新文献

Convenient production of a novel recombinant antibody against periodontitis biomarker S100A8. 便捷地制备针对牙周炎生物标志物 S100A8 的新型重组抗体。

IF 2 4区生物学

Preparative Biochemistry & Biotechnology Pub Date : 2024-11-20 DOI: 10.1080/10826068.2024.2430615

Hui-Seon Yun, Eun-Jung Kim, Byung-Gee Kim, Hee-Jin Jeong

引用次数: 0

Hybrid magnetic nanocomposites of arginine deiminase with improved stability and recyclability for biomedical applications. 具有更好稳定性和可回收性的精氨酸脱氨酶混合磁性纳米复合材料，可用于生物医学应用。

IF 2 4区生物学

Preparative Biochemistry & Biotechnology Pub Date : 2024-11-19 DOI: 10.1080/10826068.2024.2430626

Anubhuti Kawatra, Bharti Datten, Rupesh Hans, Pooja Gulati

{"title":"Hybrid magnetic nanocomposites of arginine deiminase with improved stability and recyclability for biomedical applications.","authors":"Anubhuti Kawatra, Bharti Datten, Rupesh Hans, Pooja Gulati","doi":"10.1080/10826068.2024.2430626","DOIUrl":"10.1080/10826068.2024.2430626","url":null,"abstract":"Nanocarrier-based immobilization has created new avenues for enhancing the biophysical properties of enzymes. Nanomatrices such as magnetite nanoparticles (MNPs), chitin, and chitosan with large surface areas and tunable morphology have been developed to circumvent the bottlenecks of free enzymes. The present study used MNPs to immobilize the enzyme arginine deiminase (ADI) for improved morphological control, recovery, operational stability, and easy recyclability. Hybrid magnetic arginine deiminase cross-linked enzyme aggregate (mADI-CLEA) was developed for the first time by co-aggregating ADI with magnetite nanocomposites, followed by its cross-linkage with glutaraldehyde. Structural analysis by DLS/ZETA, SEM, and FT-IR revealed their highly stable and robust nature. The resulting mADI-CLEA exhibited higher pH resistivity and thermostability than ADI-CLEA. Reusability and storage stability assay indicated that mADI-CLEA maintained more than 60% residual activity even after seven batch cycles and was stable for more than 70 days. These hybrid magnetic aggregates of ADI offer an economical and stable alternative for biomedical applications of ADI.","PeriodicalId":20401,"journal":{"name":"Preparative Biochemistry & Biotechnology","volume":" ","pages":"1-8"},"PeriodicalIF":2.0,"publicationDate":"2024-11-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142668228","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"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

Cellulase immobilization on nano-chitosan/chromium metal-organic framework hybrid matrix for efficient conversion of lignocellulosic biomass to glucose. 将纤维素酶固定在纳米壳聚糖/铬金属有机框架混合基质上，实现木质纤维素生物质到葡萄糖的高效转化。

IF 2 4区生物学

Preparative Biochemistry & Biotechnology Pub Date : 2024-11-14 DOI: 10.1080/10826068.2024.2425970

Shashi Suhag, Utkarsh Jain, Nidhi Chauhan, Vinita Hooda

{"title":"Cellulase immobilization on nano-chitosan/chromium metal-organic framework hybrid matrix for efficient conversion of lignocellulosic biomass to glucose.","authors":"Shashi Suhag, Utkarsh Jain, Nidhi Chauhan, Vinita Hooda","doi":"10.1080/10826068.2024.2425970","DOIUrl":"https://doi.org/10.1080/10826068.2024.2425970","url":null,"abstract":"In the current work, cellulase from Aspergillus niger was successfully immobilized on a novel epoxy-affixed chromium metal-organic framework/chitosan (Cr@-MIL-101/CS) support via covalent method using glutaraldehyde as a crosslinker. The bare and cellulase-bound support was characterized by using various microscopic and spectroscopic techniques. Immobilized cellulase exhibited a high immobilization yield of 0.7 ± 0.01 mg/cm2, retaining 87.5 ± 0.04% of its specific activity and displaying enhanced catalytic performance. The immobilized enzyme was maximally active at pH 5.0, temperature 65 °C and 0.9 × 10-2 mg/ml saturating substrate concentration and the half-lives of free and immobilized cellulases were approximately 9 and 19 days, respectively. The decrease in activation energy, enthalpy change, and Gibbs free energy change, coupled with an increase in entropy change upon immobilization, indicated that the enzyme's efficiency, stability, and spontaneity in catalyzing the reaction were enhanced by immobilization. Additionally, the immobilized cellulase efficiently converted rice husk cellulose to glucose, with a quantification limit of 0.05%, linear measurement ranging from 0.1 to 0.9%, and 8.5% conversion efficiency. The present method exhibited a strong correlation (R2 = 0.998) with the DNS method, validating its reliability. Notably, the epoxy/Cr@-MIL-101/CS-bound cellulase demonstrated impressive thermal and pH stabilities, retaining 50% of its activity at 75 °C and over 96% at pH levels of 4.5 and 5.0 after 12 h. Furthermore, it showed excellent reusability, preserving 80% of its activity after 15 cycles and maintaining 50% of its activity even after 20 days of storage. These results suggest that epoxy/Cr@-MIL-101/CS/cellulase composites could be very effective for large-scale cellulose hydrolysis applications.","PeriodicalId":20401,"journal":{"name":"Preparative Biochemistry & Biotechnology","volume":" ","pages":"1-21"},"PeriodicalIF":2.0,"publicationDate":"2024-11-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142626492","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"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

The effect of different light spectra on selenium bioaccumulation by Spirulina platensis cyanobacteria in flat plate photobioreactors. 平板光生物反应器中不同光谱对蓝藻硒生物累积的影响

IF 2 4区生物学

Preparative Biochemistry & Biotechnology Pub Date : 2024-11-11 DOI: 10.1080/10826068.2024.2426744

Yeganeh Shirazi, Salar Helchi, Mir Mehrshad Emamshoushtari, Sina Niakan, Elnaz Sohani, Farshid Pajoum Shariati

{"title":"The effect of different light spectra on selenium bioaccumulation by Spirulina platensis cyanobacteria in flat plate photobioreactors.","authors":"Yeganeh Shirazi, Salar Helchi, Mir Mehrshad Emamshoushtari, Sina Niakan, Elnaz Sohani, Farshid Pajoum Shariati","doi":"10.1080/10826068.2024.2426744","DOIUrl":"https://doi.org/10.1080/10826068.2024.2426744","url":null,"abstract":"Selenium (Se) plays a crucial role in human health, influencing conditions such as cancer, diabetes, and neurological disorders. With global population growth and unequal nutrient distribution threatening food security, new approaches are needed to meet the nutritional needs of the world. Se is essential for immune function, metabolism, and antioxidant defense, and in regions suffering from food insecurity and malnutrition, selenium-enriched food could offer an affordable solution. Spirulina platensis, microalgae, can bioaccumulate Se from its environment, enhancing its nutritional value. This study explores how different light spectra (red, white, yellow, and blue LEDs) affect Se bioaccumulation in Spirulina when Na2SeO3 is added to the culture medium in photobioreactors. The results show that red light made the highest Se bioaccumulation (0.118 mg.L-1), followed by white, yellow, and blue light. Se addition also increased cell dry weight by 46%, 33%, 22%, and 60%, respectively, compared to photobioreactors without Se, with biomass productivity highest under red light. Furthermore, Se boosted maximum Chl α concentration, improving photosynthetic efficiency. These findings suggest that optimizing light conditions can significantly enhance the nutritional value of Spirulina, offering a potential solution to global hunger by providing a sustainable, selenium-enriched food source.","PeriodicalId":20401,"journal":{"name":"Preparative Biochemistry & Biotechnology","volume":" ","pages":"1-11"},"PeriodicalIF":2.0,"publicationDate":"2024-11-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142626496","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"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

Optimization of microwave-assisted extraction using response surface methodology and HPLC-DAD phenolic compounds quantification from Hylocereus undatus peel and pulp cultivated in Tunisia. 利用响应面方法优化微波辅助萃取，并对突尼斯种植的 Hylocereus undatus 果皮和果肉中的 HPLC-DAD 酚类化合物进行定量。

IF 2 4区生物学

Preparative Biochemistry & Biotechnology Pub Date : 2024-11-06 DOI: 10.1080/10826068.2024.2423636

Wissal Ayouni, Hajer Riguene, Amira Yahyaoui, Marwa Dhiabi, Souad Dali, Houcine Ammar, Sirine Choura, Mohamed Chamkha, Ridha Ben Salem, Rigane Ghayth

{"title":"Optimization of microwave-assisted extraction using response surface methodology and HPLC-DAD phenolic compounds quantification from Hylocereus undatus peel and pulp cultivated in Tunisia.","authors":"Wissal Ayouni, Hajer Riguene, Amira Yahyaoui, Marwa Dhiabi, Souad Dali, Houcine Ammar, Sirine Choura, Mohamed Chamkha, Ridha Ben Salem, Rigane Ghayth","doi":"10.1080/10826068.2024.2423636","DOIUrl":"https://doi.org/10.1080/10826068.2024.2423636","url":null,"abstract":"The present study aimed to optimize the microwave-assisted extraction process for both the peel and pulp of Hylocereus undatus (white dragon fruit) cultivated in Tunisia, using response surface methodology. Total phenolic content, total flavonoid content, FRAP (ferric reducing antioxidant power), and DPPH (1,1-diphenyl-2-picrylhydrazyl) antioxidant activities were optimized. A central composite design (CCD) was applied, considering three key variables: extraction time, extraction temperature, and liquid-to-solid ratio. The optimized extraction parameters for Hylocereus undatus peel and pulp were determined respectively (9.57 min, 42.20 °C and 27.79 mL/g) and (10.08 min, 40.84 °C and 31.52 mL/g). The main phenolic compounds identified in Hylocereus undatus peel and pulp extracts using HPLC-DAD were chlorogenic and caffeic acids and rutin, quercetin, luteolin-7-O-glucoside as flavonoids. Therefore, this research has revealed the potential of a sustainable and eco-friendly process hold promise a directional option and encouraging a circular economy approach for industrial production of antioxidant-rich Hylocereus undatus extracts.","PeriodicalId":20401,"journal":{"name":"Preparative Biochemistry & Biotechnology","volume":" ","pages":"1-17"},"PeriodicalIF":2.0,"publicationDate":"2024-11-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142584000","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"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

Engineered production of 5-aminolevulinic acid in recombinant Escherichia coli BL21. 在重组大肠杆菌 BL21 中设计生产 5-氨基乙酰丙酸。

IF 2 4区生物学

Preparative Biochemistry & Biotechnology Pub Date : 2024-11-05 DOI: 10.1080/10826068.2024.2423644

Zhiwen Zhu, Bing Fu, Jiajie Lu, Peize Wang, Chuyang Yan, Fuyao Guan, Jianying Huang, Ping Yu

{"title":"Engineered production of 5-aminolevulinic acid in recombinant Escherichia coli BL21.","authors":"Zhiwen Zhu, Bing Fu, Jiajie Lu, Peize Wang, Chuyang Yan, Fuyao Guan, Jianying Huang, Ping Yu","doi":"10.1080/10826068.2024.2423644","DOIUrl":"https://doi.org/10.1080/10826068.2024.2423644","url":null,"abstract":"5-aminolevulinic acid (ALA) is a non-protein amino acid that has been widely used in the fields of medicine and agriculture. This study aims to engineer the C5 pathway of the ALA biosynthesis in Escherichia coli BL21 to enhance ALA production. The ALA synthase genes gltX, hemA, and hemL were overexpressed in E. coli BL21 to lead to the increase in the production of ALA. The sRNA RyhB was also overexpressed to downregulate the expression of ALA dehydratase to reduce the downstream bioconversion of ALA to porphobilinogen. Next, the gene arcA was knocked out by CRISPR-Cas9 technology to open the TCA cycle to promote the respiratory metabolism of the strain to reduce the feedback inhibition of heme to ALA. The fermentation conditions of the engineered strain were optimized by response surface experiments. The time-course analysis of the ALA production was carried out in a 1 L shake flask. Through these efforts, the production of ALA in engineered strain reached 2953 mg/L in a 1 L shake flask. This study contributes to the industrial production of ALA by the engineered E. coli in the future.","PeriodicalId":20401,"journal":{"name":"Preparative Biochemistry & Biotechnology","volume":" ","pages":"1-11"},"PeriodicalIF":2.0,"publicationDate":"2024-11-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142576469","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"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

Optimization of ultrasound-assisted deep eutectic solvent extraction, characterization, and bioactivities of polysaccharide from Plantaginis Semen. 超声辅助深共晶溶剂萃取的优化、表征以及植物精液多糖的生物活性。

IF 2 4区生物学

Preparative Biochemistry & Biotechnology Pub Date : 2024-11-04 DOI: 10.1080/10826068.2024.2423640

Yao Wen, Geping Chen, Lei Hu

{"title":"Optimization of ultrasound-assisted deep eutectic solvent extraction, characterization, and bioactivities of polysaccharide from Plantaginis Semen.","authors":"Yao Wen, Geping Chen, Lei Hu","doi":"10.1080/10826068.2024.2423640","DOIUrl":"https://doi.org/10.1080/10826068.2024.2423640","url":null,"abstract":"Plantaginis Semen (PS) are the dried mature seeds of Plantago asiatica L. or Plantagodepressa Willd. in the Plantago family. Its polysaccharides are important components of PS. Response surface methodology was used to optimize the ultrasonic-assisted deep eutectic solvent (DES) extraction process of PS polysaccharides (PSP). The results showed that the optimal extraction parameters were a solid-liquid ratio of 1:35 g/mL, an extraction time of 73 min, and a molar ratio of 2:1. The yield of PSP was 0.64% and 1.20% by water immersion and ultrasonic water extraction, respectively, indicating that the DES extraction method (2.21 ± 0.06%) is superior to these two methods, and the optimization effect is good. Through the α-glucosidase and α-amylase inhibition activities experiment, it was found that the IC50 values of PSPs-1 were 1122 and 220.5 μg/mL. DPPH·and ABTS+ scavenging activity experiments showed that the IC50 values of PSPs-1 were 19.2 and 4.3 μg/mL, respectively. Its molar ratio of monosaccharide composition is rhamnose: galactose: galacturonic acid: glucose: glucuronic acid: arabinose: mannose: xylose = 33.6:13.3:6.5:3:2.6:2:1.4:1. Therefore, this study can provide an experimental basis for the establishment of an industrialized production process of polysaccharides and the study of their biological activities.","PeriodicalId":20401,"journal":{"name":"Preparative Biochemistry & Biotechnology","volume":" ","pages":"1-12"},"PeriodicalIF":2.0,"publicationDate":"2024-11-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142566363","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"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

Statistical optimization and sequential scale-up of α-galactosidase production by Actinoplanes utahensis B1 from shake flask to pilot scale. 从摇瓶到中试规模，放线菌 B1 生产 α-半乳糖苷酶的统计优化和顺序放大。

IF 2 4区生物学

Preparative Biochemistry & Biotechnology Pub Date : 2024-11-01 Epub Date: 2024-05-07 DOI: 10.1080/10826068.2024.2344500

D John Babu, K Balumahendra, T C Venkateswarulu, T Sathish

{"title":"Statistical optimization and sequential scale-up of α-galactosidase production by Actinoplanes utahensis B1 from shake flask to pilot scale.","authors":"D John Babu, K Balumahendra, T C Venkateswarulu, T Sathish","doi":"10.1080/10826068.2024.2344500","DOIUrl":"10.1080/10826068.2024.2344500","url":null,"abstract":"α-Galactosidase (α-GAL) is a class of hydrolase that releases galactose from galacto-oligosaccharides and synthetic substrates such as pNPG. In this study, the production of α-GAL by Actinoplanes utahensis B1 in submerged fermentation was enhanced by using statistical methods. The effects of temperature, pH, and inoculum percentage on enzyme secretion were optimized using BBD of RSM. The optimized process was scaled up from the shake flask to the laboratory scale (5 L) and to pilot scale (30 L) using KLa based scale-up strategy. By using BBD, a maximum yield of 62.5 U/mL was obtained at a temperature of 28 °C, a pH of 6.9, and an inoculum of 6.4%. Scale-up was performed successfully and achieved a yield of 74.4 U/mL and 76.8 U/mL in laboratory scale and pilot scale fermenters. The TOST was performed to validate the scale-up strategy and the results showed a confidence level of 95% for both scales indicating the perfect execution of scale-up procedure. Through the implementation of BBD and scale-up strategy, the overall enzyme yield has been significantly increased to 76%. This is the first article to explore the scale-up of α-GAL from the A. utahensis B1 strain and provide valuable insights for industrial applications.","PeriodicalId":20401,"journal":{"name":"Preparative Biochemistry & Biotechnology","volume":" ","pages":"1216-1225"},"PeriodicalIF":2.0,"publicationDate":"2024-11-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140877104","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"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

An optimization by response surface methodology for the enhanced production of rMBSP from Pichia pastoris and study of its application. 利用响应面方法优化 Pichia pastoris 的 rMBSP 生产及其应用研究。

IF 2 4区生物学

Preparative Biochemistry & Biotechnology Pub Date : 2024-11-01 Epub Date: 2024-06-07 DOI: 10.1080/10826068.2024.2361159

Ting Li, Wenbo Li, Yao Guo, Long Han, Wen Zhang, Ronghui Liu, Hanqing Feng

{"title":"An optimization by response surface methodology for the enhanced production of rMBSP from Pichia pastoris and study of its application.","authors":"Ting Li, Wenbo Li, Yao Guo, Long Han, Wen Zhang, Ronghui Liu, Hanqing Feng","doi":"10.1080/10826068.2024.2361159","DOIUrl":"10.1080/10826068.2024.2361159","url":null,"abstract":"Background: Recombinant myofibril-bound serine proteinase (rMBSP) was successfully expressed in Pichia pastoris GS115 in our laboratory. However, low production of rMBSP in shake flask constraints further exploration of properties.Methods: A 5-L high cell density fermentation was performed and the fermentation medium was optimized. Response surface methodology (RSM) was used to optimize the culture condition through modeling three selected parameter.Results: Under the optimized culture medium (LBSM, 1% yeast powder and 1% peptone) and culture conditions (induction pH 5.5, temperature 29 °C, time 40 h), the yield of rMBSP was 420 mg/L in a 5-L fermenter, which was a 6-fold increase over thar, expressed in flask cultivation. The desired enzyme was purified by two-step, which yielded a 33.7% recovery of a product that had over 85% purity. The activity of purified rMBSP was significantly inhibited by Ca2+, Mg2+, SDS, guanidine hydrochloeide, acetone, isopropanol, chloroform, n-hexane and n-heptane. Enzymatic analysis revealed a Km of 2.89 ± 0.09 μM and a Vmax of 14.20 ± 0.12 nM•min-1 for rMBSP. LC-MS/MS analysis demonstrated the specific cleavage of bovine serum albumin by rMPSP.Conclusion: These findings suggest that rMPSP has potential as a valuable enzyme for protein science research.","PeriodicalId":20401,"journal":{"name":"Preparative Biochemistry & Biotechnology","volume":" ","pages":"1320-1328"},"PeriodicalIF":2.0,"publicationDate":"2024-11-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141284599","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"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

Kinetic and thermodynamic investigation of Rhodanese synthesized by enhanced Klebsiella oxytoca JCM 1665 strain: a comparative between the free and immobilized enzyme entrapped in alginate beads. 增强型氧合克雷伯氏菌 JCM 1665 菌株合成罗丹酶的动力学和热力学研究：游离酶与海藻酸珠子固定酶的比较。

IF 2 4区生物学

Preparative Biochemistry & Biotechnology Pub Date : 2024-11-01 Epub Date: 2024-05-02 DOI: 10.1080/10826068.2024.2347407

Babamotemi Oluwasola Itakorode, Dorcas Ibukunoluwa Itakorode, Nkem Torimiro, Raphael Emuebie Okonji

{"title":"Kinetic and thermodynamic investigation of Rhodanese synthesized by enhanced Klebsiella oxytoca JCM 1665 strain: a comparative between the free and immobilized enzyme entrapped in alginate beads.","authors":"Babamotemi Oluwasola Itakorode, Dorcas Ibukunoluwa Itakorode, Nkem Torimiro, Raphael Emuebie Okonji","doi":"10.1080/10826068.2024.2347407","DOIUrl":"10.1080/10826068.2024.2347407","url":null,"abstract":"Klebsiella oxytoca JCM 1665 was subjected to extracellular rhodanese production using a submerged fermentation technique. The organism was further engineered for higher cyanide tolerance and rhodanese yield using ethylmethanesulfonate as a mutagen. Mutagenesis resulted in an improved mutant with high cyanide tolerance (100 mM) and rhodanese yield (26.7 ± 0.67 U/mL). This yield was 4.34-fold higher than the wild strain (6.15 ± 0.65 U/mL). At temperatures ranging from 30 to 80 °C, the first-order thermal denaturation constant (Kd) for free enzyme increases from 0.00818 to 0.0333 min-1 while the immobilized enzyme increases from 0.003 to 0.0204 min-1. The equivalent half-life reduces from 99 to 21 minutes and 231 to 35 minutes, respectively. Residual activity tests were used to assess the thermodynamic parameters for both enzyme preparations. For the free enzyme, the parameters obtained were enthalpy (29.40 to 29.06 kJ.mol-1), entropy (-194.24 to -197.50 J.mol-1K-1) and Gibbs free energy (90.20 to 98.80 kJ.mol-1). In addition, for immobilized rhodanese, we obtained enthalpy (40.40 to 40.07 kJ.mol-1), entropy (-164.21 to - 165.20 J.mol-1K-1) and Gibbs free energy (91.80 to 98.40 kJ.mol-1. Regarding its operational stability, the enzyme was able to maintain 63% of its activity after being used for five cycles. Immobilized K. oxytoca rhodanese showed a marked resistance to heat inactivation compared to free enzyme forms; making it of utmost significance in many biotechnological applications.","PeriodicalId":20401,"journal":{"name":"Preparative Biochemistry & Biotechnology","volume":" ","pages":"1275-1284"},"PeriodicalIF":2.0,"publicationDate":"2024-11-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140864722","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"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