Trautwein-Schult Anke, Jankowska Dagmara, Cordes Arno, Hoferichter Petra, Klein Christina, Matros Andrea, Mock Hans-Peter, Baronian Keith, Bode Rüdiger, Kunze Gotthard
Leibniz Institute of Plant Genetics and Crop Plant Research (IPK), Gatersleben, Germany.
J Mol Microbiol Biotechnol. 2013;23(6):418-30. doi: 10.1159/000353847. Epub 2013 Sep 6.
Hyperuricemia and its symptoms are becoming increasingly common worldwide. Elevated serum uric acid levels are caused by increased uric acid synthesis from food constituents and reduced renal excretion. Treatment in most cases involves reducing alcohol intake and consumption of meat and fish or treatment with pharmaceuticals. Another approach could be to reduce uric acid level in food, either during production or consumption. This work reports the production of recombinant urate oxidase by Arxula adeninivorans and its application to reduce uric acid in a food product. The A. adeninivorans urate oxidase amino acid sequence was found to be similar to urate oxidases from other fungi (61-65% identity). In media supplemented with adenine, hypoxanthine or uric acid, induction of the urate oxidase (AUOX) gene and intracellular accumulation of urate oxidase (Auoxp) was observed. The enzyme characteristics were analyzed from isolates of the wild-type strain A. adeninivorans LS3, as well as from those of transgenic strains expressing the AUOX gene under control of the strong constitutive TEF1 promoter or the inducible AYNI1 promoter. The enzyme showed high substrate specificity for uric acid, a broad temperature and pH range, high thermostability and the ability to reduce uric acid content in food.
高尿酸血症及其症状在全球范围内正变得越来越普遍。血清尿酸水平升高是由食物成分中尿酸合成增加以及肾脏排泄减少所致。大多数情况下的治疗方法包括减少酒精摄入以及肉类和鱼类的食用量,或者进行药物治疗。另一种方法可能是在食品生产或消费过程中降低食物中的尿酸水平。这项工作报道了嗜腺嘌呤丛赤壳菌生产重组尿酸氧化酶及其在降低食品中尿酸方面的应用。发现嗜腺嘌呤丛赤壳菌尿酸氧化酶的氨基酸序列与其他真菌的尿酸氧化酶相似(同一性为61 - 65%)。在添加腺嘌呤、次黄嘌呤或尿酸的培养基中,观察到尿酸氧化酶(AUOX)基因的诱导以及尿酸氧化酶(Auoxp)的细胞内积累。从野生型菌株嗜腺嘌呤丛赤壳菌LS3的分离物以及在强组成型TEF1启动子或诱导型AYNI1启动子控制下表达AUOX基因的转基因菌株的分离物中分析了该酶的特性。该酶对尿酸表现出高底物特异性、较宽的温度和pH范围、高热稳定性以及降低食品中尿酸含量的能力。