Abstract:
:The alkane-1-monoxygenase (alkB) complex of Pseudomonas putida GPo1 has been extensively studied in the past and shown to be capable of oxidising aliphatic C(5)-C(12) alkanes to primary alcohols both in the wild-type organism by growth on C(5)-C(12) alkanes as sole carbon source and in vitro. Despite this, successful n-dodecane oxidation for the production of 1-dodecanol or dodecanoic acid has proven elusive in the past when using alkB-expressing recombinants. This article demonstrates, for the first time in vivo, by using the Escherichia coli GEC137 pGEc47ΔJ strain, that n-dodecane oxidation using this enzyme for the production of primary alcohols and carboxylic acids is feasible and in fact potentially more promising than n-octane oxidation due to lower product and substrate toxicity. Yields are reported of 1-dodecanol of up to 2 g/L(organic) and dodecanoic acid up to 19.7 g/L(organic) in a 2 L stirred tank reactor with 1L aqueous phase and 200 mL of n-dodecane as a second phase. The maximum volumetric rate of combined alcohol and acid production achieved was 1.9 g/L(organic)/h (0.35 g/L(total)/h). The maximum specific activity of combined alcohol and acid production was 7-fold lower on n-dodecane (3.5 μmol/min/g(dcw)) than on n-octane (21 μmol/min/g(dcw)); similar to the 5-fold difference observed between wild-type growth rates using the two respective alkanes as sole carbon source. Despite this, both total volumetric rate and final yield exceeded n-octane oxidation by 3.5-fold under the same conditions, due to the lower toxicity of n-dodecane and its oxidation products to E. coli compared to the 8-carbon equivalents. Substrate access limitations and the overoxidation of 1-dodecanol to dodecanoic acid were identified as the most important limitations to be addressed.
journal_name
Enzyme Microb Technoljournal_title
Enzyme and microbial technologyauthors
Grant C,Woodley JM,Baganz Fdoi
10.1016/j.enzmictec.2011.01.008subject
Has Abstractpub_date
2011-05-06 00:00:00pages
480-6issue
6-7eissn
0141-0229issn
1879-0909pii
S0141-0229(11)00027-5journal_volume
48pub_type
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更新日期:1994-06-01 00:00:00
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journal_title:Enzyme and microbial technology
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journal_title:Enzyme and microbial technology
pub_type: 杂志文章
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更新日期:2008-01-01 00:00:00
abstract::The methyl ester of 2-benzoxazolon-3-yl-acetic acid was used as an acyl donor in the penicillin amidase-catalysed transfer reaction to 7-aminodesacetoxycephalosporanic acid. The synthesis of 7-(2-benzoxazolon-3-yl-acetamido)-desacetoxycephalosporanic acid was carried out as a kinetically controlled reaction. A charact...
journal_title:Enzyme and microbial technology
pub_type: 杂志文章
doi:10.1016/0141-0229(92)90144-d
更新日期:1992-06-01 00:00:00
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journal_title:Enzyme and microbial technology
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abstract::Non-productive cellulase adsorption onto lignin is a major inhibitory mechanism preventing enzymatic hydrolysis of lignocellulosic feedstocks. Therefore, understanding of enzyme-lignin interactions is essential for the development of enzyme mixtures and processes for lignocellulose hydrolysis. We have studied cellulas...
journal_title:Enzyme and microbial technology
pub_type: 杂志文章
doi:10.1016/j.enzmictec.2013.07.003
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journal_title:Enzyme and microbial technology
pub_type: 杂志文章
doi:10.1016/j.enzmictec.2019.109440
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journal_title:Enzyme and microbial technology
pub_type: 杂志文章
doi:10.1016/j.enzmictec.2012.12.010
更新日期:2013-03-05 00:00:00
abstract::Two GH43 β-xylosidases, RS223-BX from a rice straw metagenomic library, and BoXA from Bacteroides ovatus, that share similar amino acid sequences (81% identical) and 19 of 20 active-site residues, were compared by using site-directed mutagenesis of Asp and His residues implicated in metal binding. Thus, RS223-BX is st...
journal_title:Enzyme and microbial technology
pub_type: 杂志文章
doi:10.1016/j.enzmictec.2018.03.007
更新日期:2018-07-01 00:00:00
abstract::Conventional lipase screening methods are mostly based on hydrolytic activity, which may not always be the best method to assess the enzyme activity, especially for evaluating synthetic activity. Here we developed a high throughput and visual method to screen clones with high synthetic activity and used it to assess l...
journal_title:Enzyme and microbial technology
pub_type: 杂志文章
doi:10.1016/j.enzmictec.2012.03.002
更新日期:2012-05-10 00:00:00
abstract::The characterization of the S' subsite specificity of native and ethylated alpha-chymotrypsin has been studied via acyl transfer reaction in acetonitrile containing 10 vol% of water. Using Ac-Tyr-OEt as acyl donor, we investigated the partitioning of acyl-chymotrypsins between water and amino acid and peptide-derived ...
journal_title:Enzyme and microbial technology
pub_type: 杂志文章
doi:10.1016/0141-0229(94)90125-2
更新日期:1994-07-01 00:00:00
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journal_title:Enzyme and microbial technology
pub_type: 杂志文章
doi:10.1016/j.enzmictec.2015.09.006
更新日期:2016-01-01 00:00:00
abstract::The fatty acid methyl esters and fatty acid ethyl esters are known as biodiesels which are considered to be renewable, nontoxic and biodegradable biofuels. However, the conventional biodiesels show a high crystallization temperature which is one of the most critical obstacles against the widespread biodiesel usage. Th...
journal_title:Enzyme and microbial technology
pub_type: 杂志文章
doi:10.1016/j.enzmictec.2017.03.012
更新日期:2017-07-01 00:00:00
abstract::Marine microbes have gained significant attention as potential biofactories for broad spectrum bioactive compounds. In the recent years, bioactive biosurfactants have warranted renewed interest from both environmental and clinical sectors as anti-biofilm agents due to their excellent properties of dispersing microbial...
journal_title:Enzyme and microbial technology
pub_type: 杂志文章
doi:10.1016/j.enzmictec.2018.09.008
更新日期:2019-01-01 00:00:00
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journal_title:Enzyme and microbial technology
pub_type: 杂志文章
doi:10.1016/j.enzmictec.2010.11.008
更新日期:2011-03-07 00:00:00
abstract::Enzymatic synthesis of cephalothin from 7-aminocephalosporanic acid (7-ACA) and amide derivatives of 2-thienylacetic acid (2-TA) using penicillin G acylase (pen G acylase) was studied. Two amide derivatives of 2-TA namely 2-thienylacetamide (2-TAA) and 2-thienylacetohydroxamic acid (2-TAH) were used in this study. The...
journal_title:Enzyme and microbial technology
pub_type: 杂志文章
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journal_title:Enzyme and microbial technology
pub_type: 杂志文章
doi:10.1016/j.enzmictec.2012.01.002
更新日期:2012-04-05 00:00:00
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journal_title:Enzyme and microbial technology
pub_type: 杂志文章
doi:10.1016/j.enzmictec.2015.06.002
更新日期:2015-09-01 00:00:00
abstract::beta-amylases are used in production of maltose syrup. It is shown that sweet potato beta-amylase can be purified by affinity precipitation with alginate with 80% activity yield and 44 fold purification. SDS-PAGE of the purified protein showed a single band and a subunit weight of 50 kDa. Preliminary data with soybean...
journal_title:Enzyme and microbial technology
pub_type: 杂志文章
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更新日期:2001-06-07 00:00:00
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journal_title:Enzyme and microbial technology
pub_type: 杂志文章
doi:10.1016/j.enzmictec.2014.03.006
更新日期:2014-05-10 00:00:00
abstract::The lipases from Thermomyces lanuginosus and Pseudomonas cepacia have been immobilized on octyl and cyanogen bromide (CNBr) agarose beads. The immobilization on octyl-agarose is slowed with increasing ionic strength, while the immobilization on CNBr is not significantly affected by the ionic strength. The inhibition o...
journal_title:Enzyme and microbial technology
pub_type: 杂志文章
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更新日期:2015-04-01 00:00:00
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journal_title:Enzyme and microbial technology
pub_type: 杂志文章
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更新日期:2014-06-10 00:00:00
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journal_title:Enzyme and microbial technology
pub_type: 杂志文章
doi:10.1016/j.enzmictec.2014.10.001
更新日期:2015-01-01 00:00:00
abstract::Sialic acid aldolases (E.C.4.1.3.3) catalyze the reversible aldol cleavage of N-acetyl-d-neuraminic acid (Neu5Ac) to from N-acetyl-d-mannosamine (ManNAc) and pyruvate. In this study, a sialic acid aldolase (PdNAL) from Peptoclostridium difficile NAP08 was expressed in Escherichia coli BL21 (DE3). This homotetrameric e...
journal_title:Enzyme and microbial technology
pub_type: 杂志文章
doi:10.1016/j.enzmictec.2016.07.003
更新日期:2016-10-01 00:00:00
abstract::This study examines a novel method to reduce the probability of disulfide mismatches during the refolding process by the replacement of cysteines within a protein. Specifically, Cys383 of recombinant rat procarboxypeptidase B was replaced by other amino acids to increase the refolding efficiency in vitro. Mutants C383...
journal_title:Enzyme and microbial technology
pub_type: 杂志文章
doi:10.1016/j.enzmictec.2011.04.018
更新日期:2011-07-10 00:00:00
abstract::Addition of non-ionic surfactants (NIS) is known to accelerate enzymatic lignocellulose hydrolysis. The mechanism behind this accelerating effect is still not elucidated but has been hypothesized to originate from favorable NIS-lignin interactions which alleviate non-productive adsorption of cellulases to lignin. In t...
journal_title:Enzyme and microbial technology
pub_type: 杂志文章
doi:10.1016/j.enzmictec.2011.06.015
更新日期:2011-09-10 00:00:00
abstract::A solvent-mediated method (SMM) was used to prepare supersaturated sugar solutions in hydrophobic and mixture of hydrophilic/hydrophobic ionic liquids (ILs), namely, [Bmim][Tf2N] and [Bmim][TfO]/[Bmim][Tf2N], respectively. In this method, sugars were first solubilized in a mixture of organic solvent and water (i.e. me...
journal_title:Enzyme and microbial technology
pub_type: 杂志文章
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更新日期:2019-07-01 00:00:00
abstract::Although the chaperon solvent plug was reported as a strategy to reduce aggregation before the column inlet in SEC (size-exclusion chromatography) protein refolding process, the appropriate position at which sample injected and the volume of the chaperon solvent plug have not been elucidated. Therefore, the detail of ...
journal_title:Enzyme and microbial technology
pub_type: 杂志文章
doi:10.1016/j.enzmictec.2011.05.003
更新日期:2011-07-10 00:00:00