Arce-Vázquez María Belem, Ponce-Alquicira Edith, Delgado-Fornué Ezequiel, Pedroza-Islas Ruth, Díaz-Godínez Gerardo, Soriano-Santos J
Department of Biotechnology, Metropolitan Autonomus University Mexico, Mexico.
Department of Wood, Cellulose and Paper, Biomaterials Research Center, University of Guadalajara Jalisco, Mexico.
Front Microbiol. 2016 Sep 23;7:1513. doi: 10.3389/fmicb.2016.01513. eCollection 2016.
Cyclodextrin glycosyltransferase (CGTase) is an enzyme that produces cyclodextrins (CDs) from starch and related carbohydrates, producing a mixture of α-, β-, and γ-CDs in different amounts. CGTase production, mainly by sp., depends on fermentation conditions such as pH, temperature, concentration of nutrients, carbon and nitrogen sources, among others. CGTase produces those three types of CDs, however, β-CD should prevail. Although, waxy corn starch (CS) is used industrially to obtain CGTase and CDs because of its high amylopectin content, alternative sources such as amaranth starch (AS) could be used to accomplish those purposes. AS has high susceptibility to the amylolytic activity of CGTase because of its 80% amylopectin content. Therefore, the aim of this work was evaluate the AS as carbon source for CGTase production by in a submerged fermentation. Afterwards, the CGTase was purified partially and its activity to synthesize α-, β-, and γ-CDs was evaluated using 1% AS as substrate. produced a 66 kDa CGTase (Topt = 50°C; pHopt = 8.0), from the early exponential growth phase which lasted 36 h. The maximum CGTase specific activity (106.62 ± 8.33 U/mg protein) was obtained after 36 h of culture. CGTase obtained with a Km = 0.152 mM and a Vmax = 13.4 μM/min yielded 40.47% total CDs using AS which was roughly twice as much as that of corn starch (CS; 24.48%). High costs to produce CDs in the pharmaceutical and food industries might be reduced by using AS because of its higher α-, β- and γ-CDs production (12.81, 17.94, and 9.92%, respectively) in a shorter time than that needed for CS.
环糊精糖基转移酶(CGTase)是一种能从淀粉及相关碳水化合物中产生环糊精(CDs)的酶,可产生不同比例的α-、β-和γ-环糊精混合物。CGTase的生产主要由 菌进行,这取决于发酵条件,如pH值、温度、营养物质浓度、碳源和氮源等。CGTase能产生这三种类型的环糊精,不过,β-环糊精应占主导。虽然,蜡质玉米淀粉(CS)因其高支链淀粉含量而在工业上用于生产CGTase和环糊精,但诸如苋菜籽淀粉(AS)等替代来源也可用于实现这些目的。由于AS含有80%的支链淀粉,因此它对CGTase的淀粉分解活性高度敏感。因此,本研究的目的是评估AS作为 在深层发酵中生产CGTase的碳源。之后,对CGTase进行部分纯化,并以1%的AS作为底物评估其合成α-、β-和γ-环糊精的活性。 在持续36小时的指数生长早期阶段产生了一种66 kDa的CGTase(最适温度 = 50°C;最适pH = 8.0)。培养36小时后获得了最大的CGTase比活性(106.62 ± 8.33 U/mg蛋白质)。用AS获得的CGTase的Km = 0.152 mM,Vmax = 13.4 μM/min,产生的总环糊精占40.47%,约为玉米淀粉(CS;24.48%)的两倍。由于AS在比CS所需时间更短的时间内产生更高的α-、β-和γ-环糊精产量(分别为12.81%、17.94%和9.92%),因此使用AS可能会降低制药和食品工业中环糊精的生产成本。