Hanson R L, Bembenek K S, Patel R N, Szarka L J
Department of Microbial Technology, Bristol-Myers Squibb, New Brunswick, NJ 08903.
Appl Microbiol Biotechnol. 1992 Aug;37(5):599-603. doi: 10.1007/BF00240733.
Biotransformations were developed to oxidize N epsilon-carbobenzoxy(CBZ)-L-lysine and to reduce the product keto acid to L-CBZ-oxylysine. Lysyl oxidase (L-lysine: O2 oxidoreductase, EC 1.4.3.14) from Trichoderma viride was relatively specific for L-lysine and had very low activity with N epsilon-substituted derivatives. L-Amino acid oxidase (L-amino acid: O2 oxidoreductase [deaminating], EC 1.4.3.2) from Crotalus adamanteus venom had low activity with L-lysine but high activity with N epsilon-formyl-, t-butyoxycarbonyl(BOC)-, acetyl-, trifluoroacetyl-, or CBZ-L-lysine. L-2-Hydroxyisocaproate dehydrogenase (EC 1.1.1.-) from Lactobacillus confusus catalyzed the reduction by NADH of the keto acids from N epsilon-acetyl-, trifluoroacetyl-, formyl- and CBZ-L-lysine but was inactive with the products from oxidation of L-lysine, L-lysine methyl ester, L-lysine ethyl ester or N epsilon-t-BOC-L-lysine. Providencia alcalifaciens (SC9036, ATCC 13159) was a good microbial substitute for the snake venom oxidase and also provided catalase (H2O2:H2O2 oxidoreductase EC 1.11.1.6). N epsilon-CBZ-L-Lysine was converted to CBZ-L-oxylysine in 95% yield with 98.5% optical purity by oxidation using P. alcalifaciens cells followed by reduction of the keto acid using L-2-hydroxyisocaproate dehydrogenase. NADH was regenerated using formate dehydrogenase (formate: NAD oxidoreductase, EC 1.2.1.2) from Candida boidinii. The Providencia oxidase was localized in the particulate fraction and catalase activity was predominantly in the soluble fraction of sonicated cells. The pH optima and kinetic constants were determined for the reactions.
人们开发了生物转化方法来氧化Nε-苄氧羰基（CBZ）-L-赖氨酸，并将产物酮酸还原为L-CBZ-氧基赖氨酸。来自绿色木霉的赖氨酰氧化酶（L-赖氨酸：O2氧化还原酶，EC 1.4.3.14）对L-赖氨酸具有相对特异性，而对Nε-取代衍生物的活性非常低。来自金刚王眼镜蛇毒液的L-氨基酸氧化酶（L-氨基酸：O2氧化还原酶[脱氨基]，EC 1.4.3.2）对L-赖氨酸活性低，但对Nε-甲酰基-、叔丁氧羰基（BOC）-、乙酰基-、三氟乙酰基-或CBZ-L-赖氨酸活性高。来自混淆乳杆菌的L-2-羟基异己酸脱氢酶（EC 1.1.1.-）催化NADH对Nε-乙酰基-、三氟乙酰基-、甲酰基-和CBZ-L-赖氨酸的酮酸的还原反应，但对L-赖氨酸、L-赖氨酸甲酯、L-赖氨酸乙酯或Nε-叔丁氧羰基-L-赖氨酸氧化产物无活性。产碱普罗威登斯菌（SC9036，ATCC 13159）是蛇毒氧化酶的良好微生物替代物，并且还提供过氧化氢酶（H2O2：H2O2氧化还原酶EC 1.11.1.6）。使用产碱普罗威登斯菌细胞进行氧化反应，然后使用L-2-羟基异己酸脱氢酶还原酮酸，Nε-CBZ-L-赖氨酸以95%的产率和98.5%的光学纯度转化为CBZ-L-氧基赖氨酸。使用博伊丁假丝酵母的甲酸脱氢酶（甲酸：NAD氧化还原酶，EC 1.2.1.2）再生NADH。普罗威登斯菌氧化酶定位于颗粒部分，而过氧化氢酶活性主要存在于超声处理细胞的可溶部分。测定了反应的最适pH值和动力学常数。
