Konishi Masaaki, Makino Motoki
Department of Biotechnology and Environmental Chemistry, Kitami Institute of Technology, 165 Koen-cho, Kitami, Hokkaido 090-8507, Japan.
Department of Biotechnology and Environmental Chemistry, Kitami Institute of Technology, 165 Koen-cho, Kitami, Hokkaido 090-8507, Japan.
J Biosci Bioeng. 2018 Jan;125(1):105-110. doi: 10.1016/j.jbiosc.2017.08.003. Epub 2017 Aug 26.
Mannosylerythritol lipids (MELs) are produced by several smut fungi of the Ustilaginaceae family; they are promising microbial biosurfactants and have excellent surface-active and self-assembling properties. Pseudozyma hubeiensis is a candidate for abundant MEL production and produces large amounts of 4-O-[(4'-mono-O-acetyl-2',3'-di-O-alkanoyl)-β-d-mannopyranosyl]-meso-erythritol (MEL-C). An acetyltransferase disruption mutant of P. hubeiensis, SY62-MM36, was obtained to selectively produce deacetylated 4-O-[(2',3'-di-O-alkanoyl)-β-d-mannopyranosyl]-meso-erythritol (MEL-D), and the structures of the products were determined. Lower mobility of major spots of the mutant on silica gel thin-layer chromatography verified its more hydrophilic nature than that of wild-type MEL-A, B, and C. Structural analyses confirmed the product to be MEL-D, which comprises acyl chains of caproic acid (C6:0), capric acid (C10:0), and lauric acid (C12:0). The critical micelle concentration (CMC) and the surface tension (γCMC) of the MEL-D were 2.0 × 10 M and 29.7 mN/m, respectively. SY62-MM36 also produced a minor product that was estimated as triacylated MEL-D. The triacylated MEL-D had a CMC of 3.5 × 10 M and a γCMC of 29.6 mN/m. In water, MEL-D formed a lamella liquid crystal phase over a broad range of concentrations. By fed-batch cultivation, the mutant produced 91.6 ± 6.3 g/L of MEL-D for 7 days.
甘露糖赤藓糖醇脂(MELs)由黑粉菌科的几种黑粉菌产生;它们是很有前景的微生物生物表面活性剂,具有出色的表面活性和自组装特性。湖北假丝酵母是大量生产MELs的候选菌株,可产生大量的4-O-[(4'-单-O-乙酰基-2',3'-二-O-链烷酰基)-β-D-甘露吡喃糖基]-内消旋赤藓糖醇(MEL-C)。获得了湖北假丝酵母的乙酰转移酶破坏突变体SY62-MM36,用于选择性生产脱乙酰化的4-O-[(2',3'-二-O-链烷酰基)-β-D-甘露吡喃糖基]-内消旋赤藓糖醇(MEL-D),并确定了产物的结构。突变体主要斑点在硅胶薄层色谱上的迁移率较低,证实其比野生型MEL-A、B和C具有更强的亲水性。结构分析证实产物为MEL-D,其由己酸(C6:0)、癸酸(C10:0)和月桂酸(C12:0)的酰基链组成。MEL-D的临界胶束浓度(CMC)和表面张力(γCMC)分别为2.0×10⁻⁵M和29.7mN/m。SY62-MM36还产生了一种次要产物,估计为三酰化MEL-D。三酰化MEL-D的CMC为3.5×10⁻⁶M,γCMC为29.6mN/m。在水中,MEL-D在很宽的浓度范围内形成层状液晶相。通过分批补料培养,该突变体在7天内产生了91.6±6.3g/L的MEL-D。