Dwiarti Lies, Ali Ehsan, Park Enoch Y
Laboratory of Biotechnology, Department of Applied Biological Chemistry, Faculty of Agriculture, Shizuoka University, 836 Ohya, Suruga-ku, Shizuoka 422-8529, Japan.
Bioresour Technol. 2010 Jan;101(1):14-20. doi: 10.1016/j.biortech.2009.08.032. Epub 2009 Sep 1.
This study sought to identify inhibitory factors of lipase catalyzed-fatty acid methyl esters (FAME) production from waste activated bleaching earth (wABE). During the vegetable oil refinery process, activated bleaching earth (ABE) is used for removing the impure compounds, but adsorbs vegetable oil up to 35-40% as on a weight basis, and then the wABE is discarded as waste material. The impurities were extracted from the wABE with methanol and evaluated by infra-red (IR) spectroscopy, which revealed that some were chlorophyll-plant pigments. The chlorophylls inhibited the lipase during FAME conversion from wABE. The inhibition by a mixture of chlorophyll a and b was found to be competitive. The inhibition of the enzymatic hydrolysis of waste vegetable oil contained in wABE by chlorophyll a alone was competitive, while the inhibition by chlorophyll b alone was non-competitive. Furthermore, the addition of a small amount of alkali nullified this inhibitory effect and accelerated the FAME production rate. When 0.9% KOH (w/w wABE) was added to the transesterification reaction with only 0.05% lipase (w/w wABE), the maximum FAME production rate improved 120-fold, as compared to that without the addition of KOH. The alkali-combined lipase significantly enhanced the FAME production rate from wABE, in spite of the presence of the plant pigments, and even when a lower amount of lipase was used as a catalyst.
本研究旨在确定废活性白土(wABE)中脂肪酶催化脂肪酸甲酯(FAME)生产的抑制因素。在植物油精炼过程中,活性白土(ABE)用于去除不纯化合物,但按重量计会吸附高达35 - 40%的植物油,然后wABE作为废料被丢弃。用甲醇从wABE中提取杂质,并通过红外(IR)光谱进行评估,结果表明其中一些是叶绿素类植物色素。在从wABE转化为FAME的过程中,叶绿素会抑制脂肪酶的活性。发现叶绿素a和b的混合物产生的抑制作用具有竞争性。单独叶绿素a对wABE中所含废植物油的酶促水解的抑制作用具有竞争性,而单独叶绿素b的抑制作用则是非竞争性的。此外,添加少量碱可消除这种抑制作用并加快FAME的生产速率。当在仅含有0.05%(w/w wABE)脂肪酶的酯交换反应中添加0.9%(w/w wABE)的KOH时,与不添加KOH相比,最大FAME生产速率提高了120倍。尽管存在植物色素,并且即使使用较少量的脂肪酶作为催化剂,碱结合脂肪酶仍能显著提高从wABE生产FAME的速率。