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利用统计方法高效评估和优化影响G4产生胞外木聚糖酶的培养基成分

Efficient Assessment and Optimisation of Medium Components Influencing Extracellular Xylanase Production by G4 Using Statistical Approaches.

作者信息

Ali Noor Lutphy, Foo Hooi Ling, Ramli Norhayati, Halim Murni, Thalij Karkaz M

机构信息

Department of Bioprocess Technology, Faculty of Biotechnology and Biomolecular Sciences, Universiti Putra Malaysia, Serdang 43400 UPM, Selangor, Malaysia.

Department of Medical Microbiology, College of Science, Cihan University-Erbil, Erbil 44001, Iraq.

出版信息

Int J Mol Sci. 2025 Jul 25;26(15):7219. doi: 10.3390/ijms26157219.

DOI:10.3390/ijms26157219
PMID:40806348
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC12347206/
Abstract

Xylanase is an essential industrial enzyme for degrading plant biomass, pulp and paper, textiles, bio-scouring, food, animal feed, biorefinery, chemicals, and pharmaceutical industries. Despite its significant industrial importance, the extensive application of xylanase is hampered by high production costs and concerns regarding the safety of xylanase-producing microorganisms. The utilisation of renewable polymers for enzyme production is becoming a cost-effective alternative. Among the prospective candidates, non-pathogenic lactic acid bacteria (LAB) are promising for safe and eco-friendly applications. Our investigation revealed that G4, isolated from plant sources, is a notable producer of extracellular xylanase. Improving the production of extracellular xylanase is crucial for viable industrial applications. Therefore, the current study investigated the impact of various medium components and optimised the selected medium composition for extracellular xylanase production of G4 using Plackett-Burman Design (PBD) and Central Composite Design (CCD) statistical approaches. According to BPD analysis, 8 out of the 19 investigated factors (glucose, almond shell, peanut shell, walnut shell, malt extract, xylan, urea, and magnesium sulphate) demonstrated significant positive effects on extracellular xylanase production of G4. Among them, glucose, almond shells, peanut shells, urea, and magnesium sulphate were identified as the main medium components that significantly ( < 0.05) influenced the production of extracellular xylanase of G4. The optimal concentrations of glucose, almond shells, peanut shells, urea, and magnesium sulphate, as determined via CCD, were 26.87 g/L, 16 g/L, 30 g/L, 2.85 g/L, and 0.10 g/L, respectively. The optimised concentrations resulted in extracellular xylanase activity of 2.765 U/mg, which was similar to the predicted extracellular xylanase activity of 2.737 U/mg. The CCD-optimised medium yielded a 3.13-fold enhancement in specific extracellular xylanase activity and a 7.99-fold decrease in production costs compared to the commercial de Man, Rogosa and Sharpe medium, implying that the CCD-optimised medium is a cost-effective medium for extracellular xylanase production of G4. Moreover, this study demonstrated a positive correlation between extracellular xylanase production, growth, lactic acid production and the amount of sugar utilised, implying the multifaceted interactions of the physiological variables affecting extracellular xylanase production in G4. In conclusion, statistical methods are effective in rapidly assessing and optimising the medium composition to enhance extracellular xylanase production of G4. Furthermore, the findings of this study highlighted the potential of using LAB as a cost-effective producer of extracellular xylanase enzymes using optimised renewable polymers, offering insights into the future use of LAB in producing hemicellulolytic enzymes.

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/471c/12347206/e04b1407a0a5/ijms-26-07219-g008.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/471c/12347206/3d48743bcbe9/ijms-26-07219-g002.jpg
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https://cdn.ncbi.nlm.nih.gov/pmc/blobs/471c/12347206/59b84d93a986/ijms-26-07219-g005.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/471c/12347206/0d9ff95d743c/ijms-26-07219-g006.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/471c/12347206/3833c99ef7ac/ijms-26-07219-g007.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/471c/12347206/e04b1407a0a5/ijms-26-07219-g008.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/471c/12347206/3d48743bcbe9/ijms-26-07219-g002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/471c/12347206/d2680f1461c5/ijms-26-07219-g003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/471c/12347206/59b84d93a986/ijms-26-07219-g005.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/471c/12347206/0d9ff95d743c/ijms-26-07219-g006.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/471c/12347206/3833c99ef7ac/ijms-26-07219-g007.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/471c/12347206/e04b1407a0a5/ijms-26-07219-g008.jpg
摘要

木聚糖酶是一种重要的工业酶,可用于降解植物生物质、制浆造纸、纺织、生物精练、食品、动物饲料、生物炼制、化工和制药等行业。尽管木聚糖酶具有重要的工业价值,但其广泛应用受到高生产成本以及对木聚糖酶产生菌安全性担忧的阻碍。利用可再生聚合物生产酶正成为一种具有成本效益的替代方案。在潜在的候选者中,非致病性乳酸菌有望实现安全且环保的应用。我们的研究表明,从植物源分离出的G4是一种显著的胞外木聚糖酶生产者。提高胞外木聚糖酶的产量对于可行的工业应用至关重要。因此,本研究使用Plackett-Burman设计(PBD)和中心复合设计(CCD)统计方法,研究了各种培养基成分的影响,并优化了用于G4生产胞外木聚糖酶的选定培养基组成。根据BPD分析,在19个研究因素(葡萄糖、杏仁壳、花生壳、核桃壳、麦芽提取物、木聚糖、尿素和硫酸镁)中,有8个对G4的胞外木聚糖酶生产表现出显著的正向影响。其中,葡萄糖、杏仁壳、花生壳、尿素和硫酸镁被确定为显著(<0.05)影响G4胞外木聚糖酶生产的主要培养基成分。通过CCD确定的葡萄糖、杏仁壳、花生壳、尿素和硫酸镁的最佳浓度分别为26.87 g/L、16 g/L、3O g/L、2.85 g/L和0.10 g/L。优化后的浓度导致胞外木聚糖酶活性为2.765 U/mg,与预测的胞外木聚糖酶活性2.737 U/mg相似。与商业的德氏、罗氏和夏普培养基相比,CCD优化的培养基使特定胞外木聚糖酶活性提高了3.13倍,生产成本降低了7.99倍,这意味着CCD优化的培养基是用于G4生产胞外木聚糖酶的一种具有成本效益的培养基。此外,本研究表明胞外木聚糖酶生产、生长、乳酸生产与糖利用量之间存在正相关,这意味着影响G4胞外木聚糖酶生产的生理变量之间存在多方面的相互作用。总之,统计方法对于快速评估和优化培养基组成以提高G4的胞外木聚糖酶产量是有效的。此外,本研究结果突出了利用乳酸菌作为使用优化的可再生聚合物生产胞外木聚糖酶的具有成本效益的生产者的潜力,为未来乳酸菌在生产半纤维素酶方面的应用提供了见解。

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