农用工业废料作为固态培养条件下由 Candida viswanathii 生产脂肪酶的替代品:纯化、生化特性及其用于家禽脂肪水解的潜力。

Q2 Biochemistry, Genetics and Molecular Biology Enzyme Research Pub Date : 2016-01-01 Epub Date: 2016-09-20 DOI:10.1155/2016/1353497
Alex Fernando de Almeida, Kleydiane Braga Dias, Ana Carolina Cerri da Silva, César Rafael Fanchini Terrasan, Sâmia Maria Tauk-Tornisielo, Eleonora Cano Carmona
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引用次数: 0

摘要

这项工作的目的是利用固态培养中的农用工业废料,改善粘瓦拉蒂念珠菌生产脂肪酶的条件,并纯化和评估这种酶在家禽脂肪水解中的应用。与初始条件相比,添加了 25.0%(重量比)橄榄油的混合麦麸和废大麦谷物(1:1,重量比)的脂肪酶产量提高了 322.4%。当用家禽脂肪代替橄榄油时,发现脂肪酶产量最高的是 40%(重量/重量),为 31.43 U/gds 。通过选择酵母提取物添加量(3.5%,w/w)、培养温度(30°C)和基质水分(40%,w/v),脂肪酶产量达到 157.33 U/gds。脂肪酶通过疏水相互作用色谱法纯化,经 SDS-PAGE 测定分子量为 18.5 kDa。粗制和纯化的酶分别在 pH 值为 5.0 和 50°C 以及 pH 值为 5.5 和 45°C 时显示出最佳活性。粗制脂肪酶在 50°C 时的半衰期估计为 23.5 小时,纯化脂肪酶在 40°C 时的半衰期估计为 6.7 小时。脂肪酶在许多有机溶剂中都具有高活性和稳定性。家禽脂肪在 pH 值为 4.0 时水解速度最快,初始水解速度为 33.17 mmol/L/min。因此,C. viswanathii 脂肪酶可通过经济、可持续的工艺成功生产,并可用于家禽脂肪的水解,而无需额外的酸化步骤来回收释放出的脂肪酸。
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Agroindustrial Wastes as Alternative for Lipase Production by Candida viswanathii under Solid-State Cultivation: Purification, Biochemical Properties, and Its Potential for Poultry Fat Hydrolysis.

The aims of this work were to establish improved conditions for lipase production by Candida viswanathii using agroindustrial wastes in solid-state cultivation and to purify and evaluate the application of this enzyme for poultry fat hydrolysis. Mixed wheat bran plus spent barley grain (1 : 1, w/w) supplemented with 25.0% (w/w) olive oil increased the lipase production to 322.4%, compared to the initial conditions. When olive oil was replaced by poultry fat, the highest lipase production found at 40% (w/w) was 31.43 U/gds. By selecting, yeast extract supplementation (3.5%, w/w), cultivation temperature (30°C), and substrate moisture (40%, w/v), lipase production reached 157.33 U/gds. Lipase was purified by hydrophobic interaction chromatography, presenting a molecular weight of 18.5 kDa as determined by SDS-PAGE. The crude and purified enzyme showed optimum activity at pH 5.0 and 50°C and at pH 5.5 and 45°C, respectively. The estimated half-life at 50°C was of 23.5 h for crude lipase and 6.7 h at 40°C for purified lipase. Lipase presented high activity and stability in many organic solvents. Poultry fat hydrolysis was maximum at pH 4.0, reaching initial hydrolysis rate of 33.17 mmol/L/min. Thus, C. viswanathii lipase can be successfully produced by an economic and sustainable process and advantageously applied for poultry fat hydrolysis without an additional acidification step to recover the released fatty acids.

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Enzyme Research
Enzyme Research Biochemistry, Genetics and Molecular Biology-Biochemistry
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