Substrate-constituted three-liquid-phase system: a green, highly efficient and recoverable platform for interfacial enzymatic reactions

Abstract

Highly efficient interfacial enzymatic hydrolysis of oil was achieved in a three-liquid-phase system, wherein the substrate constituted one of the phases. The enlarged interfacial area and relieved product inhibition were responsible for the high catalytic efficiency. Convenient product isolation and the high reusability of the enzyme were also demonstrated.

A Highly Efficient Three-Liquid-Phase-Based Enzymatic One-Pot Multistep Reaction System with Recoverable Enzymes for the Synthesis of Biodiesel

“…Hydrolysis of Rice Bran Oil by Lipase in Various Systems. In our previous studies, 18,25 we have successfully developed various TLPSs consisting of refined oil. However, unlike most oils, crude rice bran oil contains a large amount of amphiphilic phospholipid and FFA, which may destroy the formation of the TLPS.…”

“…The tube was vortexed for 10 s and then centrifuged at 1000g and 25 °C for 5 min. The hydrolytic activities of lipase and phospholipase in the polar phases were analyzed by the colorimetric method 18 and titration method, 24 respectively. The partition coefficient (K) and the recovery of the enzyme (Y) were determined using the following formulas…”

“…Lipase Activity Assay. The hydrolytic activity of lipase in the polar phase was analyzed by a colorimetric method, 18 which measures the rate of p-nitrophenyl octanoate (PNPC) hydrolysis. In this method, 80 μL of Tris-HCl buffer (50 mM) and 10 μL of PNPC ethanol solution (10 mM) were added into a 96-well plate and prewarmed at 37 °C for 5 min.…”

“…16,17 In our previous work, the TLPS was successfully developed and used as a single-enzymatic reaction system, which possesses certain advantages over traditional ones, such as high catalytic efficiency, good environmental compatibility, and low toxicity and energy cost. 18 Our recent finding has shown that some TLPSs exhibit high biocompatibility where two kinds of enzymes could be effectively and separately incorporated into one phase. The enzyme-enriched phase may be considered as a reusable liquid enzyme support for the EOMR, which can overcome a series of problems posed by the traditional solid coimmobilization support, such as the complex immobilization process, high immobilization cost, and difficulty in adjusting the proportion of enzymes to precisely control the reaction.…”

Correlation of three-liquid-phase equilibria involving ionic liquids

“…This process allows the recovery or separation of several compounds simultaneously, being proved as useful for metal recovery, extraction of natural products and waste water treatment [5,6]. There is also the possibility of using these systems as reaction medium, for instance in enzymatic reactions [7]. Another example of multiphase systems is liquid-liquid-liquid phase transfer catalysis.…”