title: Synthetic resin-bound truncated Candida antarctica lipase B for production of fatty acid alkyl esters by transesterification of corn and soybean oils with ethanol or butanol.
abstract: A gene encoding a synthetic truncated Candida antarctica lipase B (CALB) was generated via automated PCR and expressed in Saccharomyces cerevisiae. Western blot analysis detected five truncated CALB variants, suggesting multiple translation starts from the six in-frame ATG codons. The longest open reading frame, which corresponds to amino acids 35-317 of the mature lipase, appeared to be expressed in the greatest amount. The truncated CALB was immobilized on Sepabeads® EC-EP resin and used to produce ethyl and butyl esters from crude corn oil and refined soybean oil. The yield of ethyl esters was 4-fold greater from corn oil than from soybean oil and was 36% and 50% higher, respectively, when compared to a commercially available lipase resin (Novozym 435) using the same substrates. A 5:1 (v/v) ratio of ethanol to corn oil produced 3.7-fold and 8.4-fold greater yields than ratios of 15:1 and 30:1, respectively. With corn oil, butyl ester production was 56% higher than ethyl ester production. Addition of an ionic catalytic resin step prior to the CALB resin increased yields of ethyl esters from corn oil by 53% compared to CALB resin followed by ionic resin. The results suggest resin-bound truncated CALB has potential application in biodiesel production using biocatalysts.
authors.affiliation: United States Department of Agriculture, Agricultural Research Service, National Center for Agricultural Utilization Research, 1815 North University Street Peoria, IL 61604, United States. Stephen.Hughes@ars.usda.gov
authors.forename: Stephen R
authors.initials: SR
authors.lastname: Hughes
authors.forename: Bryan R
authors.initials: BR
authors.lastname: Moser
authors.forename: Samantha
authors.initials: S
authors.lastname: Robinson
authors.forename: Elby J
authors.initials: EJ
authors.lastname: Cox
authors.forename: Amanda J
authors.initials: AJ
authors.lastname: Harmsen
authors.forename: Jon A
authors.initials: JA
authors.lastname: Friesen
authors.forename: Kenneth M
authors.initials: KM
authors.lastname: Bischoff
authors.forename: Marjorie A
authors.initials: MA
authors.lastname: Jones
authors.forename: Rebecca
authors.initials: R
authors.lastname: Pinkelman
authors.forename: Sookie S
authors.initials: SS
authors.lastname: Bang
authors.forename: Ken
authors.initials: K
authors.lastname: Tasaki
authors.forename: Kenneth M
authors.initials: KM
authors.lastname: Doll
authors.forename: Nasib
authors.initials: N
authors.lastname: Qureshi
authors.forename: Siqing
authors.initials: S
authors.lastname: Liu
authors.forename: Badal C
authors.initials: BC
authors.lastname: Saha
authors.forename: John S
authors.initials: JS
authors.lastname: Jackson
authors.forename: Michael A
authors.initials: MA
authors.lastname: Cotta
authors.forename: Joseph O
authors.initials: JO
authors.lastname: Rich
authors.forename: Paolo
authors.initials: P
authors.lastname: Caimi
mesh_terms: D020001:1-Butanol / Q000737:chemistry / Q000378:metabolism*; D000595:Amino Acid Sequence; D001483:Base Sequence; D019149:Bioreactors; D003314:Corn Oil / Q000737:chemistry / Q000378:metabolism; D004800:Enzymes, Immobilized / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D004951:Esterification; D000431:Ethanol / Q000737:chemistry / Q000378:metabolism*; D005227:Fatty Acids / Q000737:chemistry / Q000378:metabolism*; D005656:Fungal Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D008049:Lipase / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D008969:Molecular Sequence Data; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D012117:Resins, Synthetic; D012441:Saccharomyces cerevisiae / Q000235:genetics; D013024:Soybean Oil / Q000737:chemistry / Q000378:metabolism
chemical_list: D004800:Enzymes, Immobilized; D005227:Fatty Acids; D005656:Fungal Proteins; D011994:Recombinant Proteins; D012117:Resins, Synthetic; D000431:Ethanol; D013024:Soybean Oil; D003314:Corn Oil; D020001:1-Butanol; C093667:Novozyme 435; D008049:Lipase; C431349:lipase B, Candida antarctica