Saika Azusa, Fukuoka Tokuma, Yamamoto Shuhei, Sugahara Tomohiro, Sogabe Atsushi, Morita Tomotake
Research Institute for Sustainable Chemistry, National Institute of Advanced Industrial Science and Technology (AIST), Tsukuba Central 5-2, 1-1-1 Higashi, Tsukuba, Ibaraki, 305-8565, Japan.
Toyobo Co., Ltd., Biotechnology Research Laboratory, 10-24, Toyo-cho, Tsuruga, Fukui, 914-8550, Japan.
Heliyon. 2024 Oct 28;10(21):e39789. doi: 10.1016/j.heliyon.2024.e39789. eCollection 2024 Nov 15.
Mannosylerythritol lipids (MELs) are glycolipid biosurfactants produced by various yeasts. MEL producers produce mainly di-acylated MELs (consisting of two fatty acid chains). Among them, is a di-acylated MEL-B (d-MEL-B) producer. In a previous study, we generated an acyltransferase-deleted strain of . (Δ), which selectively produced mono-acylated MEL-D (m-MEL-D, consisting of one fatty acid chain), but not d-MEL-B. However, m-MEL-D productivity in Δ was low, and oil consumption was significantly reduced compared to the parent strain. Based on these findings, we hypothesized that the d-MEL-B produced by the parent strain may act as an emulsifier in the culture medium, leading to easier utilization of the oil. By contrast, the m-MEL-D produced by Δ may not have the ability to emulsify oil, thus the oil is used inefficiently and productivity of m-MEL-D is low. Therefore, we expected that adding d-MEL-B to the culture medium during Δ cultivation would increase m-MEL-D production. To enhance the oil consumption and m-MEL-D production of Δ, d-MEL-B and chemical surfactants were added to the culture medium as emulsifiers during Δ cultivation. Adding d-MEL-B enhanced both the oil consumption and m-MEL-D production of Δ; Tween 20 and Triton X-100 also showed enhancement effects. As expected, d-MEL-B, Tween20 and TritonX-100, showed marked olive oil emulsification activity, whereas m-MEL-D did not. These results strongly support our hypothesis and significantly improve m-MEL-D productivity.
甘露糖赤藓糖醇脂(MELs)是由多种酵母产生的糖脂类生物表面活性剂。MELs产生菌主要产生二酰化MELs(由两条脂肪酸链组成)。其中,[具体菌株名称]是二酰化MEL - B(d - MEL - B)产生菌。在之前的一项研究中,我们构建了[具体菌株名称]的酰基转移酶缺失菌株(Δ[具体菌株名称]),该菌株选择性地产生单酰化MEL - D(m - MEL - D,由一条脂肪酸链组成),而不产生d - MEL - B。然而,Δ[具体菌株名称]中m - MEL - D的产量较低,与亲本菌株相比,油脂消耗显著降低。基于这些发现,我们推测亲本菌株产生的d - MEL - B可能在培养基中充当乳化剂,从而使油脂更易被利用。相比之下,Δ[具体菌株名称]产生的m - MEL - D可能没有乳化油脂的能力,因此油脂利用效率低下,m - MEL - D的产量也较低。因此,我们期望在Δ[具体菌株名称]培养过程中向培养基中添加d - MEL - B会增加m - MEL - D的产量。为了提高Δ[具体菌株名称]的油脂消耗和m - MEL - D产量,在Δ[具体菌株名称]培养过程中,将d - MEL - B和化学表面活性剂作为乳化剂添加到培养基中。添加d - MEL - B提高了Δ[具体菌株名称]的油脂消耗和m - MEL - D产量;吐温20和曲拉通X - 100也显示出增强作用。正如预期的那样,d - MEL - B、吐温20和曲拉通X - 100表现出显著的橄榄油乳化活性,而m - MEL - D则没有。这些结果有力地支持了我们的假设,并显著提高了m - MEL - D的产量。