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从本地品种的牙髓中提取酪氨酸酶的纯化、特性分析、固定化,及其在 l-3,4-二羟苯丙氨酸批量生产中的应用。

Tyrosinase from the pulps of local cultivars of Purification, characterization, immobilization, and application in the batch production of l-3,4-dihydroxyphenylalanine.

机构信息

Department of Biochemistry, Olabisi Onabanjo University, Ago-Iwoye, Nigeria.

Department of Physical and Chemical Sciences, Elizade University, Ilara-Mokin, Nigeria.

出版信息

Prep Biochem Biotechnol. 2024 Sep;54(8):1098-1105. doi: 10.1080/10826068.2024.2324084. Epub 2024 Mar 6.

Abstract

Tyrosinase, an enzyme involved in browning reactions in plants/crops exposed to mechanical injury, was isolated from the pulp of some different locally available bananas (, , and ). Tyrosinase from the pulps was extracted, purified, immobilized, and characterized. Thereafter, the potentials of the immobilized tyrosinase in the possible production of l-3,4-dihydroxyphenylalanine (L-DOPA) in an improvised batch reactor was exploited using tyrosine and ascorbate as the substrates. L-DOPA production was monitored via thin-layer chromatography and spectrophotometry (Arnow's method). L-DOPA is a drug that is used in the treatment of Parkinson's disease. Hence, this study exploited a non-chemical route for its synthesis using the tyrosinase obtained from the banana pulps. The purified tyrosinase had an optimum pH and temperature of 6.5 and 7.0, respectively. The molecular weight of the purified tyrosinase was 45 kDa. Quercetin and resorcinol both competitively inhibited the purified tyrosinase from the three cultivars. Immobilized tyrosinase produced the highest concentration (0.60 mM) of L-DOPA after 8 h in an improvised batch reactor. The tyrosinase in the banana pulps serves as a cheap and readily available green route for the possible production of L-DOPA.

摘要

多酚氧化酶,一种参与植物/作物机械损伤后褐变反应的酶,从一些不同的当地香蕉(、、和)的果肉中分离出来。从果肉中提取、纯化、固定化和表征多酚氧化酶。此后,利用酪氨酸和抗坏血酸作为底物,在一个临时分批反应器中探索了固定化多酚氧化酶在可能生产左旋多巴(L-DOPA)中的潜力。通过薄层层析和分光光度法(Arnow 法)监测 L-DOPA 的生产。L-DOPA 是一种用于治疗帕金森病的药物。因此,本研究利用从香蕉果肉中获得的多酚氧化酶,通过非化学途径合成 L-DOPA。纯化的多酚氧化酶的最适 pH 和温度分别为 6.5 和 7.0。纯化的多酚氧化酶的分子量为 45 kDa。槲皮素和间苯二酚均竞争性抑制三种品种的纯化多酚氧化酶。固定化多酚氧化酶在临时分批反应器中 8 小时后产生了最高浓度(0.60 mM)的 L-DOPA。香蕉果肉中的多酚氧化酶是一种廉价且易得的绿色途径,可用于可能生产 L-DOPA。

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