用酵母和转化酶研究甜菜糖蜜中蔗糖水解的抑制因素

Catalysts Pub Date : 2024-05-17 DOI:10.3390/catal14050330
Mikael Sjölin, Maria Djärf, Mohamed Ismail, Herje Schagerlöf, Ola Wallberg, Rajni Hatti-Kaul, Mahmoud Sayed
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引用次数: 0

摘要

甜菜糖蜜是制糖业的一种低价值副产品。它含有大量蔗糖(约 50%(w/w)),可用于多种不同用途,例如作为生产燃料(乙醇)和生物基化学品(如 5-羟甲基糠醛 (HMF))的原料。为了生产平台化学品,蔗糖被水解成单体 C6 糖:葡萄糖和果糖。在相同的蔗糖浓度(300 克/升)和工艺条件下,比较糖蜜和纯蔗糖溶液的水解速率,发现其特定反应速率要慢得多(纯蔗糖和粗糖蜜的 Qp/x,60min = 93 和 70 gprod L-1 h-1 gcell-1)。为了弄清糖蜜抑制酶水解速率的原因,研究了糖蜜的粘度、无机和有机成分的影响。此外,还测试了糖蜜和处理过的糖蜜对纯酶--转化酶(来自酿酒酵母,0.05 毫克/毫升)--的影响,并与使用贝克酵母全细胞(3 毫克/毫升)进行的水解进行了比较。结果表明,钾具有抑制作用(Qp/x,60min = 76 gprod L-1 h-1 gcell-1),一般在盐浓度较高时(Qp/x,60min = 67 gprod L-1 h-1 gcell-1),这可能与溶液的盐浓度较高有关,也可能与不同离子的协同效应有关,当使用的离子浓度是糖蜜浓度的四倍时。此外,粘度和蔗糖纯度似乎也有影响,纯蔗糖溶液和来自糖厂的浓汁的水解率(Qp/x,60min = 97 gprod L-1 h-1 gcell-1)高于粘度较高的糖蜜型溶液(Qp/x,60min = 70-74 gprod L-1 h-1 gcell-1)。为了进一步了解不同成分对转化酶活性的影响,我们进行了一项硅学调查,结果表明高浓度盐会影响蔗糖与酶活性位点的结合,从而导致反应速率降低。这方面的知识对于今后扩大水解工艺的规模非常重要,因为水解率降低需要更大的体积才能提供一定的生产率,这就需要更大的工艺设备,从而增加了投资成本。
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Investigating the Inhibitory Factors of Sucrose Hydrolysis in Sugar Beet Molasses with Yeast and Invertase
Sugar beet molasses is a low-value byproduct from the sugar industry. It contains significant amounts of sucrose (approx. 50% (w/w)), which can be used for many different applications, for example, as feedstock for the production of fuel (as ethanol) and biobased chemicals such as 5-hydoxymethyl furfural (HMF). To produce platform chemicals, sucrose is hydrolyzed into its monomeric C6 sugars: glucose and fructose. When comparing the hydrolysis rates of molasses with a pure sucrose solution, the specific reaction rate is much slower (Qp/x,60min = 93 and 70 gprod L−1 h−1 gcell−1 for pure sucrose and crude molasses, respectively) at the same sucrose concentration (300 g/L) and process conditions. To clarify why molasses inhibits the enzymatic hydrolysis rate, the influence of its viscosity and inorganic and organic composition was investigated. Also, the effects of molasses and treated molasses on pure enzymes, invertase (from Saccharomyces cerevisiae, 0.05 mg/mL), compared with hydrolysis using whole cells of Baker’s yeast (3 mg/mL), were tested. The results indicate an inhibitory effect of potassium (Qp/x,60min = 76 gprod L−1 h−1 gcell−1), generally at high salt concentrations (Qp/x,60min = 67 gprod L−1 h−1 gcell−1), which could be correlated to the solution’s high salt concentrations and possibly the synergistic effects of different ions when applying concentrations that were four times that in the molasses. Also, the viscosity and sucrose purity seem to have an effect, where pure sucrose solutions and thick juice from the sugar mill yielded higher hydrolysis rates (Qp/x,60min = 97 gprod L−1 h−1 gcell−1) than molasses-type solutions with a higher viscosity (Qp/x,60min = 70–74 gprod L−1 h−1 gcell−1). Attempting to further understand the effects of different components on the invertase activity, an in silico investigation was performed, indicating that high salt concentrations affected the binding of sucrose to the active site of the enzyme, which can result in a lower reaction rate. This knowledge is important for future scale-up of the hydrolysis process, since reduced hydrolysis rates require larger volumes to provide a certain productivity, requiring larger process equipment and thereby higher investment costs.
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