Enzyme and Microbial Technology最新文献

{"title":"Dual antioxidant and antibacterial potential of β-cryptoxanthin and a novel biosurfactant from Kocuria marina DAGII","authors":"Daiji Brahma ,&nbsp;Anwesha Mahanty ,&nbsp;Prity Chatterjee ,&nbsp;Riya Dey ,&nbsp;Abheerup Sarker ,&nbsp;Sagnik Kulavi ,&nbsp;Debjani Dutta","doi":"10.1016/j.enzmictec.2026.110821","DOIUrl":"10.1016/j.enzmictec.2026.110821","url":null,"abstract":"<div><div>In the present study, the combinatory role of beta-cryptoxanthin (β-CRX) and novel biosurfactant (BS) produced by <em>Kocuria marina</em> DAGII was explored. Spectral analysis of biosurfactant showed <em>a</em> strong peak at 265 nm. Characterization of biosurfactant was performed using CHNSO (carbon, hydrogen, nitrogen, sulphur, and oxygen) and MALDI-TOF (matrix-assisted laser desorption ionization-time of flight) analysis. MALDI-TOF spectra showed an intense peak at 379.7 <em>m/z</em>, indicating the presence of structural components like sugars, lipids, and clusters of fragment ions closely related to rhamnolipid (glycolipid) like biosurfactant. CHNSO analysis of partially purified biosurfactant revealed the presence of nitrogen (4.675 %), carbon (30.815 %), hydrogen (7.417 %), and oxygen (25.428 %). A combination of BS and β-CRX demonstrated higher antioxidant activity compared to BS alone. The highest inhibition against <em>Staphylococcus epidermidis</em> MTCC 3615 and <em>Pseudomonas aeruginosa</em> MTCC 741 was observed for the combined effect of BS (32 mg/ml) and β-CRX (0.1 mg/ml). However, the lowest antibacterial activity was observed against <em>Staphylococcus aureus</em> MTCC 96 (0.3 ± 0.041 cm), likely due to its strong biofilm formation. Membrane integrity and permeability assays revealed enhanced activity against Gram-positive <em>Staphylococcus epidermidis</em> having OD₂₆₀ value 1.23 ± 0.060 and OD₄₂₀value 0.157 ± 0.01. The increased outer membrane permeability was assessed through the NPN uptake assay in <em>Pseudomonas aeruginosa</em>. Antibiofilm assay showed good antibiofilm activity (66.14 ± 0.13 %) against <em>Staphylococcus epidermidis</em> for combined effect of BS (32 mg/ml) and β-CRX (0.1 mg/ml). Overall, the study suggests that a combination of BS and β-CRX has promising potential as a dual antioxidant and antibacterial agent, particularly effective against these tested opportunistic skin pathogens.</div></div>","PeriodicalId":11770,"journal":{"name":"Enzyme and Microbial Technology","volume":"196 ","pages":"Article 110821"},"PeriodicalIF":3.7,"publicationDate":"2026-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146071142","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":"Structural, quantitative, and functional characterization of N-glycans in porcine pancreatin extract, lipase, and α-amylase","authors":"Chulmin Moon,&nbsp;Chang Myeong Jeong,&nbsp;Chi Soo Park,&nbsp;Han Seul Lee,&nbsp;Kyuran Kim,&nbsp;Haeun Byeon,&nbsp;Daeun Eom,&nbsp;Siwon Kim,&nbsp;Seojeong Lee,&nbsp;Ha Hyung Kim","doi":"10.1016/j.enzmictec.2026.110830","DOIUrl":"10.1016/j.enzmictec.2026.110830","url":null,"abstract":"<div><div>Porcine pancreatin extract (PPE), an enzyme mixture comprising porcine pancreatic lipase (PPL), porcine pancreatic α-amylase (PPA), and proteases, facilitates the digestion of fats, carbohydrates, and proteins and is widely used to treat pancreatic insufficiency. <em>N</em>-glycosylation is a critical post-translational modification influencing enzyme stability and activity. This study presents the <em>N</em>-glycan profiling of PPE, PPL, and PPA to evaluate their structural roles in enzymatic function. <em>N</em>-glycans were enzymatically released, procainamide-labeled, and analyzed using liquid chromatography–tandem mass spectrometry. Relative quantity of each <em>N</em>-glycan was expressed as a percentage of total <em>N</em>-glycan peak areas, and absolute quantity (pmol) of total <em>N</em>-glycans was calculated using calibration curves. We identified 32 (PPE), 31 (PPL), and 10 (PPA) <em>N</em>-glycans exhibiting diverse structural features, including sialylation and fucosylation. Sialylation was more abundant in PPE (25.6 %) and PPL (29.3 %) than in PPA (10.9 %), whereas fucosylation was prevalent in 67.4 %, 66.9 %, and 54.3 % of structures, respectively. Absolute quantity of total <em>N</em>-glycans was 422 pmol/mg PPE (1.01 pmol/total USP unit), 602 pmol/mg PPL (3.13 × 10⁻² pmol/pmol), and 131 pmol/mg PPA (0.72 × 10⁻² pmol/pmol). Functional assays after deglycosylation, desialylation, and defucosylation revealed that sialylation and fucosylation influenced the activities of PPE and PPL but had minimal effects on PPA. These results indicate that sialylation and fucosylation are critical for PPE and PPL function but not for PPA. This is the first <em>N</em>-glycan profiling of PPE, PPL, and PPA, and demonstrates the relevance of <em>N</em>-glycans in maintaining structural stability and regulating the enzymatic activity of PPE and PPL.</div></div>","PeriodicalId":11770,"journal":{"name":"Enzyme and Microbial Technology","volume":"196 ","pages":"Article 110830"},"PeriodicalIF":3.7,"publicationDate":"2026-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146225967","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":"The underlying thermal inactivation mechanism and corresponding effective stabilization strategy for Rhizopus oryzae lipase","authors":"Huanhuan Li ,&nbsp;Yao Wang ,&nbsp;Kun Wang ,&nbsp;Li Xu ,&nbsp;Wenyue Liu ,&nbsp;Lingfang Gu ,&nbsp;Xiaoman Xie ,&nbsp;Yunjun Yan","doi":"10.1016/j.enzmictec.2026.110822","DOIUrl":"10.1016/j.enzmictec.2026.110822","url":null,"abstract":"<div><div>Poor thermal stability of <em>Rhizopus oryzae</em> lipase (ROL) has long limited its industrial applicability. In this study, we systematically investigated the underlying mechanism for its inactivation and correspondingly developed an effective stabilization strategy. Kinetic analyses revealed that ROL thermal inactivation process followed a two-step model, demonstrating its inactivation proceeds a distinct intermediate state. Various spectroscopic characterizations further suggested that activity loss was closely associated with the progressive unfolding of the enzyme, as evidenced by the disruption of secondary structures and a pronounced increase in fluorescence intensity resulting from the exposure of hydrophobic clusters. To enhance its thermostability, based on the above-discovered mechanism, a compound stabilizer system was developed and further optimized via a combination of Plackett-Burman design and response surface methodology. The obtained optimal formula comprised of 35.94 % (m/v) glucose, 34.82 % (m/v) sorbitol, and 2.50 mol/L NaCl, which could effectively preserve 97.7 % of the initial activity after 3 h of incubation at 60 °C, in contrast to only 29.5 % residual activity of the control. Thus, this study elucidated the underlying thermal inactivation mechanism for ROL and developed a practical and efficient stabilization strategy with potential prospect for industrial application.</div></div>","PeriodicalId":11770,"journal":{"name":"Enzyme and Microbial Technology","volume":"196 ","pages":"Article 110822"},"PeriodicalIF":3.7,"publicationDate":"2026-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146112385","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":"Structure-guided engineering of CYP82D213 for enhanced triptonide biosynthesis","authors":"Shijun Yuan ,&nbsp;Lingfang Feng ,&nbsp;Yao Xu ,&nbsp;Meng Xia ,&nbsp;Ping Su ,&nbsp;Yating Hu ,&nbsp;Yifeng Zhang ,&nbsp;Luqi Huang","doi":"10.1016/j.enzmictec.2026.110832","DOIUrl":"10.1016/j.enzmictec.2026.110832","url":null,"abstract":"<div><div>Triptonide (TN) is a highly oxidized, bioactive diterpenoid triepoxide from <em>Tripterygium wilfordii</em> with promising pharmaceutical applications. While the early steps of its biosynthesis have been elucidated, the mechanism responsible for the critical terminal tri-epoxidation remains to be elucidated. Here, to address the limitation posed by the low intrinsic activity of CYP82D213, we employed a comprehensive protein engineering strategy. Structure-guided mutagenesis pinpointed the key residues responsible for substrate recognition (G128, W129) and dynamic conformation (L396). Motif-driven engineering yielded the synergistic double mutant H425Q/L459M, which enhanced activity 2.30-fold. Subsequent surface charge optimization, further improved performance. The combination of these beneficial mutations in the triple mutant H425Q/L459M/T365R resulted in a 2.63-fold increase in TN production compared to the wild-type and revealed a putative dynamic catalytic mechanism that may involve substrate repositioning. These results provide a rational design strategy for complex multi-step enzyme catalysis.</div></div>","PeriodicalId":11770,"journal":{"name":"Enzyme and Microbial Technology","volume":"196 ","pages":"Article 110832"},"PeriodicalIF":3.7,"publicationDate":"2026-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146226014","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":"Harnessing seaweed-associated marine Streptomyces sp. OSs-1 for nanotechnology-driven cost-effective enzyme production using agricultural waste","authors":"Vaishali R. Majithiya,&nbsp;Sangeeta D. Gohel","doi":"10.1016/j.enzmictec.2026.110831","DOIUrl":"10.1016/j.enzmictec.2026.110831","url":null,"abstract":"<div><div>Protease and amylase are industrially significant enzymes whose production must balance purity, productivity, and cost-efficiency. This study investigates the isolation, phenotypic and genetic characterization, growth kinetics, immobilization, and statistical optimization of enzyme co-production by <em>Streptomyces</em> sp. OSs-1, associated with the red seaweed <em>Sarconema</em> sp., isolated from the intertidal zone of Okha, Gujarat. The red alga <em>Sarconema</em> is industrially valuable due to its carrageenan production and interesting association with bioactive microbial communities. A Plackett-Burman design identified five significant variables influencing enzyme production, which were further optimized using RSM with a Box-Behnken design. ANOVA confirmed that NaCl concentration and incubation time were the most critical factors (p &lt; 0.05). Under optimal conditions (3 % NaCl, 5-day incubation), maximum amylase (375.91 ± 23.70 U/mL) and protease (294.89 ± 15.07 U/mL) production was achieved. Graphical tools, including Pareto charts and interaction plots, validated these outcomes. To reduce production costs, agricultural wastes were explored as alternative substrates. Sorghum straw supported the highest enzyme co-production (amylase: 206.87 ± 7.07 U/mL, protease: 315.08 ± 14.14 U/mL), along with the highest cell mass (0.0043 g/mL) and specific growth rate (0.0062 h⁻¹). Enzyme immobilization using sodium alginate, zinc oxide nanoparticles, and hydroxyapatite enhanced enzyme stability and reusability. The highest immobilized activities were obtained with sodium alginate+ 5 % glutaraldehyde for amylase (316.39 U/g), and with sodium alginate+zinc oxide nanoparticles+hydroxyapatite+ 10 % glutaraldehyde for protease (298.65 U/g). Immobilized enzymes showed improved thermal and pH stability. This study demonstrates that incorporating agricultural waste substrates, advanced immobilization techniques, and nanotechnology offers a cost-effective and scalable solution for industrial enzyme production from seaweed-associated <em>Streptomyces</em> sp. OSs-1.</div></div>","PeriodicalId":11770,"journal":{"name":"Enzyme and Microbial Technology","volume":"196 ","pages":"Article 110831"},"PeriodicalIF":3.7,"publicationDate":"2026-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146171626","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":"The potential and innovative applications of CRISPR gene editing technology in enzyme gene development","authors":"Youmin Zhu","doi":"10.1016/j.enzmictec.2025.110799","DOIUrl":"10.1016/j.enzmictec.2025.110799","url":null,"abstract":"<div><div>The CRISPR gene editing technology is simple in design and highly efficient, making it the most widely used gene editing tool today. At present, CRISPR gene editing technology has shown a certain application value in enzyme development, but its application potential has not been fully developed. CRISPR gene editing technology can not only be used to knockin enzyme genes and knockout genes that are not conducive to enzyme expression, but can also be applied to single-base editing of enzyme genes, tandem sgRNA for multi-enzyme gene editing, sgRNA library for enzyme screening, endogenous enzyme gene modification, transcriptional activation or inhibition of enzyme gene expression, and fluorescence imaging of enzyme genes. Especially, this review innovatively proposes for the first time that CRISPR gene editing technology can be used for site specific fusion of enzyme genes, cell surface display of endogenous enzymes, and knockin of super long DNA for simultaneous expression of multiple enzymes, providing new ideas for maximizing the value of CRISPR gene editing technology in enzyme development in the future.</div></div>","PeriodicalId":11770,"journal":{"name":"Enzyme and Microbial Technology","volume":"195 ","pages":"Article 110799"},"PeriodicalIF":3.7,"publicationDate":"2026-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145827035","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":"Efficient production of active recombinant cholesterol oxidase from Rhodococcus erythropolis in Escherichia coli via his-tag assisted refolding strategy for cholesterol oxidation","authors":"Meka Saima Perdani ,&nbsp;Dwini Normayulisa Putri ,&nbsp;Dita Ariyanti ,&nbsp;Cintiya Septa Hasannah ,&nbsp;Fitri Yuliasari ,&nbsp;Ibnu Maulana Hidayatullah ,&nbsp;Heri Hermansyah ,&nbsp;Yosuke Fukutani ,&nbsp;Masafumi Yohda","doi":"10.1016/j.enzmictec.2025.110805","DOIUrl":"10.1016/j.enzmictec.2025.110805","url":null,"abstract":"<div><div>This study reports the cloning, expression, and characterization of recombinant cholesterol oxidase (ChOx) from Rhodococcus erythropolis in Escherichia coli BL21(DE3). The ChOx gene was inserted into the pET23b(+) vector using primers designed with site-specific modifications that enabled His-tag activation, expression, and purification by nickel-affinity chromatography to yield a protein of ∼55 kDa. A urea-gradient dialysis refolding protocol was employed to recover active enzyme from inclusion bodies. The purified enzyme exhibited a specific activity of 13.68 U/mg, representing a 2.5-fold improvement over previously reported recombinant ChOx yields with a recovery yield of 76.16 %. Cholesterol oxidation assays optimized enzyme concentration, substrate concentration, and temperature. Optimum activity occurred at 0.4 mg/mL enzyme and 20 mM cholesterol, achieving &gt; 80 % substrate conversion, while the enzyme retained stability up to 50 °C. This work demonstrates a His-tag-assisted refolding strategy that efficiently produces active recombinant Rhodococcus erythropolis ChOx (RhoChOx). The optimization of catalytic parameters highlights the enzyme’s robustness under diverse conditions. These findings establish RhoChOx as a promising biocatalyst for industrial cholesterol oxidation and provide a methodological framework applicable to other recombinant oxidases.</div></div>","PeriodicalId":11770,"journal":{"name":"Enzyme and Microbial Technology","volume":"195 ","pages":"Article 110805"},"PeriodicalIF":3.7,"publicationDate":"2026-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145838175","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":"Enzymatic synthesis of sucrose esters: Advances and challenges in high-efficiency and regioselective catalysis","authors":"Lei Zhao ,&nbsp;Jinghui Lai ,&nbsp;Shenghui Zhang ,&nbsp;Mengqi Dai ,&nbsp;Enxiang Zong ,&nbsp;Youqiang Xu ,&nbsp;Xiuting Li","doi":"10.1016/j.enzmictec.2025.110806","DOIUrl":"10.1016/j.enzmictec.2025.110806","url":null,"abstract":"<div><div>As an eco-friendly and non-toxic nonionic surfactant, enzymatically synthesized sucrose esters (SEs) are increasingly valued across the food, cosmetic, and pharmaceutical industries due to their environmentally benign production characteristics. However, current enzymatic synthesis faces several fundamental constraints, like limited diversity of enzyme resources restricting process versatility, insufficient catalytic activity and thermostability of enzymes leading to suboptimal reaction efficiency, and prohibitive enzyme production costs hindering industrial scalability. To address these critical challenges, this review presents a systematic summarize of SEs synthetic technologies and their emerging applications, with particular emphasis on elucidating enzymatic synthetic mechanisms. Due to limited enzyme resources, we innovatively propose a high-throughput bioinformatics-driven enzyme mining platform that employs SEs synthase sequences as molecular probes for screening the SEs synthases, identifying previously uncharacterized enzyme candidates with potentially enhanced catalytic efficiencies. Through systematic bottleneck analysis and a forward-looking perspective, our work establishes a theoretical foundation for developing cost-effective enzymatic processes while providing a valuable repository of potential enzyme candidates for the first time. These findings offer useful guidelines for promoting enzymatic SEs production.</div></div>","PeriodicalId":11770,"journal":{"name":"Enzyme and Microbial Technology","volume":"195 ","pages":"Article 110806"},"PeriodicalIF":3.7,"publicationDate":"2026-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145881383","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":"Levansucrase from Gluconacetobacter diazotrophicus. Key residues involved in levan synthesis","authors":"Ana G. Martínez ,&nbsp;Yamira Quintero ,&nbsp;Duniesky Martínez ,&nbsp;Alexis Musacchio ,&nbsp;Odet Céspedes ,&nbsp;Carmen Menéndez","doi":"10.1016/j.enzmictec.2025.110802","DOIUrl":"10.1016/j.enzmictec.2025.110802","url":null,"abstract":"<div><div>Levansucrase from <em>Gluconacetobacter diazotrophicus</em> (LsdA, EC 2.4.1.10) produces short-chain fructooligosaccharides, mainly 1-kestotriose, and levan polysaccharide from sucrose. By random mutagenesis, twenty-five LsdA residues were identified as being crucial for levan synthesis, with the substitution of these residues resulting in a decrease or complete elimination of levan synthesis. Fourteen residues (R171, H172, S228, R229, T243, F304, N306, E314, E327, R334, A369, D398, H419, and G432) were identified within the active site cavity, while eleven were dispersed across the protein. Saturation mutagenesis of H172 and R171 (-1 subsite) showed that the H172E/P mutant primarily exhibited sucrose hydrolysis, while the H172S and R171K-H172S variants were less affected in transfructosylation. Furthermore, HPAEC-PAD analysis revealed that the H172S and R171K-H172S variants synthesized 1-kestotriose, 6-kestotriose, 6G-kestotriose, and 1,1-kestotetraose. These variants had specific activity values similar to those of the native LsdA. However, the synthesis of 1,6-kestotetraose was found to be compromised, indicating a loss of the processive mechanism, suggesting that these variants have lost the ability to elongate via β-(2→6) links.</div></div>","PeriodicalId":11770,"journal":{"name":"Enzyme and Microbial Technology","volume":"195 ","pages":"Article 110802"},"PeriodicalIF":3.7,"publicationDate":"2026-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145753851","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":"Immobilization of α-glucosidase on polystyrene plates: A practical application to α-amylase detection","authors":"Jihye Jung ,&nbsp;Verónica Mora-Sanz ,&nbsp;Nerea Briz ,&nbsp;Bernd Nidetzky","doi":"10.1016/j.enzmictec.2026.110818","DOIUrl":"10.1016/j.enzmictec.2026.110818","url":null,"abstract":"<div><div>α-Glucosidase is in high demand for applications involving the digestion of α-configured glycoside and saccharide substrates. Areas of application include industrial processing in food/feed and chemistry, as well as medicine and analytics. With an analytical application for assaying α-amylase in mind, we conducted a study on the immobilization of α-glucosidases from <em>Bacillus stearothermophilus</em> and <em>Saccharomyces cerevisiae</em> on support materials based on polystyrene and polymethacrylate. The carriers had nine different surface functional groups: none (plain material), carboxylate, sulfate, thiol, hydroxy, amine, sulfonyl chloride, epoxy, and PEG300. Based on preliminary results obtained with polymer beads, we selected the α-glucosidase from <em>B. stearothermophilus</em> and polystyrene plates with surface functional groups for detailed studies on enzyme immobilization. Carrier activity (<em>A</em>_c), immobilized enzyme effectiveness (ƞ), and reusability were evaluated depending on protein loading. Sulfonyl chloride (-SO₂Cl) showed efficient and stable immobilization (<em>A</em>_c up to 279 U/m² carrier; ƞ 58 %; <em>A</em>_c after 6 uses, 171 U/m² carrier). The α-glucosidase immobilized onto sulfonyl chloride-polystyrene carrier was used in an assay of α-amylase activity as a representative analytical application. 4-Nitrophenyl 4,6-ethylidene-α-<span>D</span>-maltoheptaoside was used as the chromogenic substrate in the α-amylase assay. The results indicated that the immobilized α-glucosidase was suitable for detecting α-amylase concentrations ranging from 0.1 µg/mL to 0.5 µg/mL (0.3 – 1.3 U/L as observed activities). Additionally, the results can be readily applied in a microtiter plate-based assay. Collectively, our results reveal the basic requirements for immobilizing α-glucosidase on polystyrene for the advancement of analytical assays.</div></div>","PeriodicalId":11770,"journal":{"name":"Enzyme and Microbial Technology","volume":"195 ","pages":"Article 110818"},"PeriodicalIF":3.7,"publicationDate":"2026-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145972933","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}