Department of Chemistry, GC University Lahore, Katchehry Road, Lahore, 54600 Pakistan.
Biotechnol Appl Biochem. 2010 Mar 12;55(3):131-7. doi: 10.1042/BA20090248.
The inductive effect of L-ascorbate on the microbiological production of L-dopa (3,4-dihydroxy-L-phenylalanine) from Aspergillus oryzae NRRL-1560 was investigated. All biochemical reactions were performed aerobically using mould mycelia as a source of enzyme tyrosinase and acetate buffer (pH 3.0) as an extractant. L-Tyrosine as a substrate was added at a level of 2.5 mg/ml. Maximal L-dopa production (1.876 mg/ml) was achieved when L-ascorbate (5.0 mg/ml) was added 6 min after the initiation of the biochemical reaction at 50 degrees C, consuming 2.144 mg/ml L-tyrosine. The performance of fuzzy-logic control of the reaction was found to be highly promising for improvement of the substrate conversion rate (~80%). After optimizing the reaction conditions, particularly the addition of L-ascorbate, an increase in L-dopa yield of 22.96% was achieved compared with the control (without ascorbate addition) when the process variables, namely buffer pH, L-tyrosine concentration and reaction temperature, were further identified using a two-factorial Plackett-Burman design.
考察了 L-抗坏血酸对米曲霉 NRRL-1560 微生物生产 L-多巴(3,4-二羟基-L-苯丙氨酸)的诱导作用。所有生化反应均在有氧条件下进行,使用霉菌菌丝作为酶酪氨酸酶的来源,醋酸盐缓冲液(pH 3.0)作为提取物。以 2.5mg/ml 的浓度添加 L-酪氨酸作为底物。当在 50°C 下开始生化反应 6 分钟后添加 5.0mg/ml 的 L-抗坏血酸时,可获得最大的 L-多巴产量(1.876mg/ml),消耗 2.144mg/ml 的 L-酪氨酸。发现模糊逻辑控制反应的性能非常有希望提高底物转化率(~80%)。在优化反应条件后,特别是添加 L-抗坏血酸后,与对照(不添加抗坏血酸)相比,当进一步使用两因素 Plackett-Burman 设计确定过程变量(即缓冲液 pH 值、L-酪氨酸浓度和反应温度)时,L-多巴的产量增加了 22.96%。