Purification and Immobilization of a Novel Enantioselective Lipase from Tsukamurella tyrosinosolvents for Efficient Resolution of Ethyl 2-(2-oxopyrrolidin-1-yl) Butyrate

Abstract

A highly enantioselective lipase from Tsukamurella tyrosinosolvents E105 was purified via ultrasonic extraction, precipitation, and chromatographic steps. The enzyme was purified about 38-fold with the recovery yield of 9 % and was confirmed as a dimer protein consisting of two identical subunits with a molecular mass of 24 kDa. The purified lipase was used to catalyze resolution of racemic ethyl 2-(2-oxopyrrolidin-1-yl) butyrate to (S)-2-(2-oxopyrrolidin-1-yl) butyric acid. The maximum activity of such lipase was obtained at pH 7.5, 35 °C, and the highest relative activity (156.80 %) was observed in the presence of 0.5 mM Co. Subsequently, the lipase was encapsulated within a mixture of 3 % sodium alginate and 0.8 % carrageenan, and then cross-linked with 0.6 % glutaraldehyde to enhance its biocatalytic capability and stability. Comparing with 36.9 % product yield and 97.5 % product ee of free lipase, the highest product yield of 46.3 % and ee of 98.5 % for immobilized lipase were achieved with the presence of 20 mM substrate. In addition, the reusability of immobilized lipase was also investigated, which could maintain 63.7 % of its initial conversion yield after seven repeated batch reactions. Thus, the evaluated enantioselective lipase in this work has a good potential for further industrial application.

Enantioselective Conversion of Racemic Sotalol to R(-)-Sotalol by Lipase AP6

“…This resulted in high yield (42 %) and enantiomeric purity (71.7 %) of (R)-enantiomer of racemic (R,S)-1-phenylethanol. The results were also in line with the results of Huang et al [41] , who reported that the activity and enantioselectivity of lipase from Tsukamurella tyrosinosolvents E105 is enhanced by immobilization and is resulted in improved yield of (S)-2-(2oxopyrrolidin-1-yl)butyric acid after enantioselective resolution of racemic ethyl 2-(2-oxopyrrolidin-1-yl) butyrate. Chromatograms of sotalol after incubation with free and immobilized-lipase AP6 were shown in fig.…”

“…When compared to the reaction catalysed by free-enzyme, the immobilized-enzyme catalysed study has yielded good enantiomeric purity at each time point. This was because immobilization will improve the stability, enantioselectivity and catalytic activity of the enzyme [37,40,41] .…”

Dynamics of PHA-Accumulating Bacterial Communities Fed with Lipid-Rich Liquid Effluents from Fish-Canning Industries

“…Considering the use of a lipid-rich waste as a substrate for the PHAs accumulation, it is necessary for the hydrolysis of triglycerides into free fatty acids before their subsequent transformation to PHAs [ 98 ]. The bacteria that were potentially able to accumulate PHAs with a lipolytic activity were Acidovorax [ 99 ], Acinetobacter [ 100 ], Azospirillum [ 101 ], Chryseobacterium [ 102 ], Comamonas [ 103 ], Corynebacterium [ 104 ], Devosia [ 105 ], Dokdonella [ 106 ], Dyella [ 107 ], Ferruginibacter [ 108 ], Flavobacterium [ 109 ], Gemmobacter [ 77 ], Gordonia [ 110 ], Leifsonia [ 111 ], Lysobacter [ 112 ], Niveispirillum [ 113 ], Novosphingobium [ 114 ], Pandoraea [ 115 ], Polynucleobacter [ 83 ], Pseudomonas [ 98 ], Qipengyuania [ 116 ], Rhizobium [ 117 ], Stenotrophomonas [ 118 ], Taibaiella [ 119 ], Thermomonas [ 120 ], and Tsukamurella [ 121 ].…”

Identification of a Phosphorus-Solubilizing Tsukamurella Tyrosinosolvens Strain and its Effect on the Bacterial Diversity of the Rhizosphere Soil of Peanuts Growth-Promoting

“…It was early reported that strains of this genus were opportunistic pathogens (Bouza et al 2009), but recent studies have shown that human infections with Tsukamurella are seldom because the genus is a kind of saprophyte bacterium (Safaei et al 2017). The functions of the Tsukamurella are diverse, especially T. tyrosinosolvens is extensively researched, this strain was able to use alkanes as carbon sources (Romanova et al 2019), produce the biosurfactants of oligosaccharide lipids (Vollbrecht et al 1998), oxidize CO (Park et al 2009), degrade the dibutyl phthalate and hexadecane (Ogawa et al 2009), and produce lipase to resolve ethyl 2-(2-oxopyrrolidin-1-yl) butyrate (Huang et al 2016). The above showed that T. tyrosinosolvens is mostly related to the degradation and metabolism of xenobiotics, but the impact on plants is only seen in Sang's report ( 2013), which T. tyrosinosolvens isolated from pepper roots could against Phytophthora capsici and reduce anthracnose occurrence under the field conditions.…”