Enzyme and Microbial Technology最新文献_第2页

Structural, quantitative, and functional characterization of N-glycans in porcine pancreatin extract, lipase, and α-amylase 猪胰蛋白酶提取物、脂肪酶和α-淀粉酶中n -聚糖的结构、定量和功能表征。

IF 3.7 3区生物学 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY

Enzyme and Microbial Technology

Pub Date : 2026-05-01 Epub Date: 2026-02-12 DOI: 10.1016/j.enzmictec.2026.110830

Chulmin Moon, Chang Myeong Jeong, Chi Soo Park, Han Seul Lee, Kyuran Kim, Haeun Byeon, Daeun Eom, Siwon Kim, Seojeong Lee, Ha Hyung Kim

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. N-glycosylation is a critical post-translational modification influencing enzyme stability and activity. This study presents the N-glycan profiling of PPE, PPL, and PPA to evaluate their structural roles in enzymatic function. N-glycans were enzymatically released, procainamide-labeled, and analyzed using liquid chromatography–tandem mass spectrometry. Relative quantity of each N-glycan was expressed as a percentage of total N-glycan peak areas, and absolute quantity (pmol) of total N-glycans was calculated using calibration curves. We identified 32 (PPE), 31 (PPL), and 10 (PPA) N-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 N-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 N-glycan profiling of PPE, PPL, and PPA, and demonstrates the relevance of N-glycans in maintaining structural stability and regulating the enzymatic activity of PPE and PPL.

猪胰酶提取物（PPE）是一种由猪胰脂肪酶（PPL）、猪胰α-淀粉酶（PPA）和蛋白酶组成的酶混合物，促进脂肪、碳水化合物和蛋白质的消化，被广泛用于治疗胰腺功能不全。n -糖基化是影响酶稳定性和活性的关键翻译后修饰。本研究介绍了PPE、PPL和PPA的n -聚糖谱，以评估它们在酶功能中的结构作用。n -聚糖酶解，普鲁卡因酰胺标记，并使用液相色谱-串联质谱分析。每个n -聚糖的相对数量以占总n -聚糖峰面积的百分比表示，并通过校准曲线计算总n -聚糖的绝对数量（pmol）。我们鉴定了32个（PPE）、31个（PPL）和10个（PPA） n -聚糖，它们具有不同的结构特征，包括唾液化和聚焦化。唾液酰化在PPE（25.6 %）和PPL（29.3 %）中比在PPA（10.9 %）中更为丰富，而聚焦化在67.4 %、66.9 %和54.3 %的结构中普遍存在。总n -聚糖的绝对数量分别为422 pmol/mg PPE （1.01 pmol/总USP单位）、602 pmol/mg PPL（3.13 × 10-2 pmol/pmol）和131 pmol/mg PPA（0.72 × 10-2 pmol/pmol）。去糖基化、去脂酰化和去聚焦化后的功能分析显示，唾液酰化和聚焦化影响PPE和PPL的活性，但对PPA的影响很小。这些结果表明，唾液化和聚焦化对PPE和PPL的功能至关重要，但对PPA不起作用。这是PPE、PPL和PPA的第一个n -聚糖分析，并证明了n -聚糖在维持PPE和PPL的结构稳定性和调节酶活性方面的相关性。

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引用次数: 0

The underlying thermal inactivation mechanism and corresponding effective stabilization strategy for Rhizopus oryzae lipase 米根霉脂肪酶热失活机制及有效稳定策略研究。

IF 3.7 3区生物学 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY

Enzyme and Microbial Technology

Pub Date : 2026-05-01 Epub Date: 2026-01-30 DOI: 10.1016/j.enzmictec.2026.110822

Huanhuan Li , Yao Wang , Kun Wang , Li Xu , Wenyue Liu , Lingfang Gu , Xiaoman Xie , Yunjun Yan

Poor thermal stability of Rhizopus oryzae 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.

米根霉脂肪酶（Rhizopus oryzae lipase， ROL）热稳定性差，长期限制了其工业应用。在这项研究中，我们系统地研究了其失活的潜在机制，并相应地制定了有效的稳定策略。动力学分析表明，ROL热失活过程遵循两步模型，表明其失活过程处于一个明显的中间状态。各种光谱表征进一步表明，活性丧失与酶的逐渐展开密切相关，二级结构的破坏和暴露于疏水团簇导致的荧光强度的显著增加证明了这一点。为了提高其热稳定性，基于上述机理，开发了复合稳定剂体系，并结合Plackett-Burman设计和响应面法对其进行了进一步优化。得到的最佳配方为35.94 % （m/v）葡萄糖、34.82 % （m/v）山梨醇、2.50 mol/L NaCl，经60℃培养3 h后，可有效保持97.7 %的初始活性，而对照的剩余活性仅为29.5 %。因此，本研究阐明了ROL潜在的热失活机制，并开发了一种实用高效的稳定策略，具有潜在的工业应用前景。

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引用次数: 0

Structure-guided engineering of CYP82D213 for enhanced triptonide biosynthesis 结构导向工程CYP82D213增强雷公藤甲素的生物合成。

IF 3.7 3区生物学 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY

Enzyme and Microbial Technology

Pub Date : 2026-05-01 Epub Date: 2026-02-14 DOI: 10.1016/j.enzmictec.2026.110832

Shijun Yuan , Lingfang Feng , Yao Xu , Meng Xia , Ping Su , Yating Hu , Yifeng Zhang , Luqi Huang

Triptonide (TN) is a highly oxidized, bioactive diterpenoid triepoxide from Tripterygium wilfordii 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.

雷公藤内酯是雷公藤中一种高度氧化、具有生物活性的三氧化二萜类化合物，具有广阔的医药应用前景。虽然其生物合成的早期步骤已经阐明，但负责关键末端三环氧化的机制仍有待阐明。在这里，为了解决CYP82D213固有活性低所带来的限制，我们采用了一种综合的蛋白质工程策略。结构导向诱变确定了负责底物识别（G128, W129）和动态构象（L396）的关键残基。基序驱动工程获得了增效双突变体H425Q/L459M，其活性增强了2.3倍。后续的表面电荷优化，进一步提高了性能。在H425Q/L459M/T365R三重突变体中，这些有益突变的组合导致TN产量比野生型增加2.63倍，并揭示了可能涉及底物重新定位的假定动态催化机制。这些结果为复杂的多步骤酶催化提供了合理的设计策略。

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引用次数: 0

Harnessing seaweed-associated marine Streptomyces sp. OSs-1 for nanotechnology-driven cost-effective enzyme production using agricultural waste 利用与海藻相关的海洋链霉菌sp. OSs-1利用农业废弃物进行纳米技术驱动的低成本酶生产

IF 3.7 3区生物学 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY

Enzyme and Microbial Technology

Pub Date : 2026-05-01 Epub Date: 2026-02-12 DOI: 10.1016/j.enzmictec.2026.110831

Vaishali R. Majithiya, Sangeeta D. Gohel

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 Streptomyces sp. OSs-1, associated with the red seaweed Sarconema sp., isolated from the intertidal zone of Okha, Gujarat. The red alga Sarconema 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 < 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 Streptomyces sp. OSs-1.

蛋白酶和淀粉酶是工业上重要的酶，其生产必须平衡纯度、生产率和成本效益。研究了古吉拉特邦奥卡潮间带红藻Streptomyces sp. ss -1的分离、表型和遗传特征、生长动力学、固定化和酶产酶统计优化。红藻Sarconema是工业上有价值的，因为它的卡拉胶生产和有趣的联系与生物活性微生物群落。Plackett-Burman设计确定了影响酶产量的五个显著变量，并使用Box-Behnken设计进一步优化了RSM。方差分析证实NaCl浓度和孵育时间是最关键的影响因素（p <； 0.05）。在最佳条件下（3 % NaCl，培养5 d），淀粉酶（375.91 ± 23.70 U/mL）和蛋白酶（294.89 ± 15.07 U/mL）产量最高。图形工具，包括帕累托图和交互图，验证了这些结果。为了降低生产成本，研究了农业废弃物作为替代基质。高粱秸秆的酶协同产量最高（淀粉酶：206.87 ± 7.07 U/mL，蛋白酶：315.08 ± 14.14 U/mL），细胞质量最高（0.0043 g/mL），特定生长速率最高（0.0062 h−1）。酶固定化使用海藻酸钠、氧化锌纳米颗粒和羟基磷灰石增强酶的稳定性和可重用性。海藻酸钠+ 5 %戊二醛对淀粉酶的固定化活性最高（316.39 U/g），海藻酸钠+氧化锌纳米颗粒+羟基磷灰石+ 10 %戊二醛对蛋白酶的固定化活性最高（298.65 U/g）。固定化酶表现出更好的热稳定性和pH稳定性。该研究表明，结合农业废弃物底物、先进的固定化技术和纳米技术，为从海藻相关链霉菌sp. OSs-1中生产工业酶提供了一种具有成本效益和可扩展的解决方案。

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引用次数: 0

The potential and innovative applications of CRISPR gene editing technology in enzyme gene development CRISPR基因编辑技术在酶基因开发中的潜力和创新应用。

IF 3.7 3区生物学 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY

Enzyme and Microbial Technology

Pub Date : 2026-04-01 Epub Date: 2025-12-20 DOI: 10.1016/j.enzmictec.2025.110799

Youmin Zhu

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.

CRISPR基因编辑技术设计简单，效率高，是当今应用最广泛的基因编辑工具。目前，CRISPR基因编辑技术在酶的开发中已显示出一定的应用价值，但其应用潜力尚未得到充分开发。CRISPR基因编辑技术不仅可以用于敲入酶基因和敲除不利于酶表达的基因，还可以用于酶基因的单碱基编辑、串联sgRNA用于多酶基因编辑、sgRNA文库用于酶筛选、内源性酶基因修饰、酶基因表达的转录激活或抑制、酶基因的荧光成像等。特别是，本文创新性地首次提出了CRISPR基因编辑技术可用于酶基因的位点特异性融合、内源性酶的细胞表面展示、超长DNA的敲入以同时表达多种酶，为未来最大化CRISPR基因编辑技术在酶开发中的价值提供了新的思路。

引用次数: 0

Efficient production of active recombinant cholesterol oxidase from Rhodococcus erythropolis in Escherichia coli via his-tag assisted refolding strategy for cholesterol oxidation 利用his标签辅助胆固醇氧化的重折叠策略在大肠杆菌中高效生产重组红红球菌活性胆固醇氧化酶

IF 3.7 3区生物学 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY

Enzyme and Microbial Technology

Pub Date : 2026-04-01 Epub Date: 2025-12-24 DOI: 10.1016/j.enzmictec.2025.110805

Meka Saima Perdani , Dwini Normayulisa Putri , Dita Ariyanti , Cintiya Septa Hasannah , Fitri Yuliasari , Ibnu Maulana Hidayatullah , Heri Hermansyah , Yosuke Fukutani , Masafumi Yohda

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 > 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.

本研究报道了红红红球菌重组胆固醇氧化酶（ChOx）在大肠杆菌BL21（DE3）中的克隆、表达和特性分析。将ChOx基因插入pET23b（+）载体，使用经过位点特异性修饰设计的引物，通过镍亲和层析使his标签激活、表达和纯化，产生约55 kDa的蛋白。采用尿素梯度透析复折叠方法从包涵体中回收活性酶。纯化酶的比活性为13.68 U/mg，比先前报道的重组ChOx产量提高2.5倍，回收率为76.16 %。胆固醇氧化试验优化酶浓度、底物浓度和温度。酶的最佳活性为0.4 mg/mL和20 mM胆固醇，达到>； 80 %底物转化率，酶在50°C下保持稳定性。这项工作证明了his标签辅助的重折叠策略，有效地产生活性重组红polis红球菌ChOx （RhoChOx）。催化参数的优化突出了酶在不同条件下的鲁棒性。这些发现确立了RhoChOx作为工业胆固醇氧化生物催化剂的前景，并为其他重组氧化酶提供了适用的方法框架。

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引用次数: 0

Enzymatic synthesis of sucrose esters: Advances and challenges in high-efficiency and regioselective catalysis 酶法合成蔗糖酯：高效和区域选择性催化的进展和挑战

IF 3.7 3区生物学 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY

Enzyme and Microbial Technology

Pub Date : 2026-04-01 Epub Date: 2025-12-26 DOI: 10.1016/j.enzmictec.2025.110806

Lei Zhao , Jinghui Lai , Shenghui Zhang , Mengqi Dai , Enxiang Zong , Youqiang Xu , Xiuting Li

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.

酶促合成蔗糖酯（SEs）作为一种环保无毒的非离子表面活性剂，因其对环境无害的生产特性，在食品、化妆品和制药等行业越来越受到重视。然而，目前的酶合成面临着几个基本的限制，如酶资源的有限多样性限制了工艺的通用性，酶的催化活性和热稳定性不足导致反应效率不理想，酶的生产成本过高阻碍了工业可扩展性。为了解决这些关键的挑战，本文综述了SEs合成技术及其新兴应用的系统总结，特别强调阐明酶合成机制。由于酶资源有限，我们创新地提出了一种高通量生物信息学驱动的酶挖掘平台，该平台采用SEs合酶序列作为分子探针筛选SEs合酶，识别以前未表征的具有潜在增强催化效率的候选酶。通过系统的瓶颈分析和前瞻性的观点，我们的工作为开发具有成本效益的酶促工艺奠定了理论基础，同时首次提供了有价值的潜在候选酶库。这些发现为促进酶促SEs的产生提供了有用的指导。

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引用次数: 0

Levansucrase from Gluconacetobacter diazotrophicus. Key residues involved in levan synthesis 重氮养糖醋杆菌的左旋蔗糖酶。利凡合成中涉及的关键残基

IF 3.7 3区生物学 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY

Enzyme and Microbial Technology

Pub Date : 2026-04-01 Epub Date: 2025-12-12 DOI: 10.1016/j.enzmictec.2025.110802

Ana G. Martínez , Yamira Quintero , Duniesky Martínez , Alexis Musacchio , Odet Céspedes , Carmen Menéndez

Levansucrase from Gluconacetobacter diazotrophicus (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.

重氮营养葡萄糖醋杆菌（glucconacetobacter diazotrophicus, LsdA, EC 2.4.1.10）的左旋蔗糖酶（Levansucrase）产生短链低聚果糖，主要是1-酮三糖，并从蔗糖中产生左旋多糖。通过随机诱变，鉴定出25个对levan合成至关重要的LsdA残基，这些残基的替代导致levan合成的减少或完全消除。在活性位点空腔内鉴定出14个残基（R171、H172、S228、R229、T243、F304、N306、E314、E327、R334、A369、D398、H419和G432）， 11个残基分散在整个蛋白中。H172和R171（-1亚位）的饱和诱变表明，H172E/P突变体主要表现为蔗糖水解，而H172S和R171K-H172S突变体在转果糖基化方面受影响较小。此外，HPAEC-PAD分析显示，H172S和R171K-H172S变体合成1-酮三糖、6-酮三糖、6g -酮三糖和1,1-酮四糖。这些变体具有与本地LsdA相似的特定活性值。然而，1,6-酮四糖的合成被发现受到损害，表明过程机制的丧失，这表明这些变体已经失去了通过β-（2→6）连接延长的能力。

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引用次数: 0

Immobilization of α-glucosidase on polystyrene plates: A practical application to α-amylase detection α-葡萄糖苷酶在聚苯乙烯板上的固定化：α-淀粉酶检测的实际应用

IF 3.7 3区生物学 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY

Enzyme and Microbial Technology

Pub Date : 2026-04-01 Epub Date: 2026-01-11 DOI: 10.1016/j.enzmictec.2026.110818

Jihye Jung , Verónica Mora-Sanz , Nerea Briz , Bernd Nidetzky

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 Bacillus stearothermophilus and Saccharomyces cerevisiae 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 B. stearothermophilus and polystyrene plates with surface functional groups for detailed studies on enzyme immobilization. Carrier activity (A_c), immobilized enzyme effectiveness (ƞ), and reusability were evaluated depending on protein loading. Sulfonyl chloride (-SO₂Cl) showed efficient and stable immobilization (A_c up to 279 U/m² carrier; ƞ 58 %; A_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-D-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.

α-葡萄糖苷酶在α-配位糖苷和糖底物的消化方面有很高的应用需求。应用领域包括食品/饲料和化学的工业加工，以及医学和分析。以α-淀粉酶为研究对象，研究了嗜热脂肪芽孢杆菌和酿酒酵母α-葡萄糖苷酶在聚苯乙烯和聚甲基丙烯酸酯载体材料上的固定化。载体具有9种不同的表面官能团：无（普通材料）、羧酸盐、硫酸盐、硫醇、羟基、胺、磺酰氯、环氧树脂和PEG300。基于聚合物微球的初步结果，我们选择了嗜热脂芽孢杆菌α-葡萄糖苷酶和带有表面官能团的聚苯乙烯板进行酶固定的详细研究。载体活性（Ac）、固定化酶效率（）和可重复使用性根据蛋白质负载进行评估。磺酰氯（-SO2Cl）表现出高效稳定的固定化效果（Ac可达279 U/m2载体；±58 %；6次后Ac为171 U/m2载体）。将α-葡萄糖苷酶固定在磺酰氯-聚苯乙烯载体上，作为α-淀粉酶活性测定的代表性应用。α-淀粉酶的显色底物为4-硝基苯基4,6-乙基-α- d -麦芽糖苷。结果表明，固定化α-葡萄糖苷酶适用于检测α-淀粉酶浓度范围为0.1 ~ 0.5 µg/mL（观察活性为0.3 ~ 1.3 U/L）。此外，结果可以很容易地应用于微量滴度板为基础的分析。总的来说，我们的结果揭示了在聚苯乙烯上固定α-葡萄糖苷酶的基本要求，以促进分析分析的发展。

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引用次数: 0

Corrigendum to “Effects of overexpression of the human intestinal alkaline phosphatase gene on the expression of related genes in intestinal epithelium-like cells” [Enzyme Microb. Technol. 195 (2026) 110807] 《人肠道碱性磷酸酶基因过表达对肠上皮样细胞相关基因表达的影响》的勘误[酶微生物]。[j].中国机械工程，2015,(5):391 - 391。

IF 3.7 3区生物学 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY

Enzyme and Microbial Technology

Pub Date : 2026-04-01 Epub Date: 2026-01-23 DOI: 10.1016/j.enzmictec.2026.110820

Seiko Noda , Shiho Ishii , Asako Yamada , Sadako Matsui , Hideo Orimo , Masae Goseki-Sone

引用次数: 0