Production of 12-hydroxy dodecanoic acid methyl ester using a signal peptide sequence-optimized transporter AlkL and a novel monooxygenase

The extractives of C. camphora old bark were used to cure some special diseases; however, the biomedical constituents of extractives of C. camphora old bark are still not completely explained. Therefore, 350 °C-based pyrolysis- GC/MS technology was used to identify the top value-added biomedical constituents of old bark-based benzene/ethanol extractives from C. camphora trunk. 74 chemical constituents representing 99.17% were identified from 86 peaks. The main components are as: Dodecanoic acid, 1-(hydroxymethyl)-1,2-ethanediyl ester (15.717%), Octacosanoic acid, methyl ester (10.357%), Abietic acid (8.483%), Heptacosane (5.678%), Ethyl alcohol (5.282%), Hexadecanoic acid, 2-hydroxy-, methyl ester (3.365%), Hexacosanoic acid, methyl ester (2.936%), 1,2-Benzenedicarboxylic acid, butyl 2-methylpropyl ester (2.928%), 2-Methoxy-4-vinylphenol (2.692%), Hexadecanoic acid, ethyl ester (2.312%), Hexacosane (2.076%), Tetracosanoic acid, methyl ester (2.013%), etc. The analytical result showed that the 350 °C pyrolyzate of benzene/ethanol extractives from C. camphora old bark is abundant in biomedical constituents, and also contains other bioactive components, which can be used as top value-added materials of high-grade cosmetic, food, spice and chemical solvents.

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GC-MS Analysis of Active and Applicable Compounds in Methanol Extract of Sweet Star Fruit (Averrhoa carambola L.) Leaves

HAYATI Journal of Biosciences ◽

10.4308/hjb.28.1.12 ◽

2021 ◽

Vol 28 (1) ◽

pp. 12

Author(s):

Ni Putu Adriani Astiti ◽

Yan Ramona

Keyword(s):

Methyl Ester ◽

Methanol Extract ◽

Acid Methyl Ester ◽

Acid Ethyl Ester ◽

Methanol Solution ◽

Dodecanoic Acid ◽

Octadecanoic Acid ◽

Hexadecanoic Acid ◽

Averrhoa Carambola ◽

Acid Methyl

The leaves of star fruit plants (Averrhoa carambola) have been traditionally used to cure many diseases, such as hypertension and fever. Besides, the leaves of this plant have also widely been used as the main raw material in lawar (a Balinese traditional food) making. In this research, the methanol extract of this plant leaves was analyzed and identified for active compound contents. The main objective of this research was to investigate types of possible active and applicable compounds contained in such leaves, previously extracted/macerated in methanol solution. Analysis was conducted by applying GC-MS instrumentation using methanol solution. The results showed that 10 possible active and applicable compounds (Butane, 1,1-diethoxy-3 methyl-(CAS)1.1-DII, Dodecanoic acid, methyk ester(CAS) methyl, Dodecanoic acid, methyl ester (CAS) Ethyl Laun, Pentadecanoic acid ethyl ester, Hexadecanoic acid methyl ester (CAS) Methyl pa, OCTADECA 9.12 DIENOIC ACID METHYL, 9-Octadecenoic acid methyl ester (E)-(CAS), Octadecanoic acid methyl ester, and (E) 9-Octadecanoic acid ethyl ester) were identified in methanol extract of such leaves. Among those compounds, Butane, 1,1-diethoxy-3-methyl and Hexadecanoic acid, methyl ester was two most abundance constituents with percentage of peak areas of 35.67% and 26.93%, respectively.

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Outer Membrane Protein AlkL Boosts Biocatalytic Oxyfunctionalization of Hydrophobic Substrates in Escherichia coli

Applied and Environmental Microbiology ◽

10.1128/aem.00949-12 ◽

2012 ◽

Vol 78 (16) ◽

pp. 5724-5733 ◽

Cited By ~ 76

Author(s):

Mattijs K. Julsing ◽

Manfred Schrewe ◽

Sjef Cornelissen ◽

Inna Hermann ◽

Andreas Schmid ◽

...

Keyword(s):

Escherichia Coli ◽

Methyl Ester ◽

Membrane Protein ◽

Outer Membrane ◽

Outer Membrane Protein ◽

Acid Methyl Ester ◽

Dodecanoic Acid ◽

Content Type ◽

E Coli ◽

Acid Methyl

ABSTRACTThe outer membrane of microbial cells forms an effective barrier for hydrophobic compounds, potentially causing an uptake limitation for hydrophobic substrates. Low bioconversion activities (1.9 U gcdw−1) have been observed for the ω-oxyfunctionalization of dodecanoic acid methyl ester by recombinantEscherichia colicontaining the alkane monooxygenase AlkBGT ofPseudomonas putidaGPo1. Using fatty acid methyl ester oxygenation as the model reaction, this study investigated strategies to improve bacterial uptake of hydrophobic substrates. Admixture of surfactants and cosolvents to improve substrate solubilization did not result in increased oxygenation rates. Addition of EDTA increased the initial dodecanoic acid methyl ester oxygenation activity 2.8-fold. The use of recombinantPseudomonas fluorescensCHA0 instead ofE. coliresulted in a similar activity increase. However, substrate mass transfer into cells was still found to be limiting. Remarkably, the coexpression of thealkLgene ofP. putidaGPo1 encoding an outer membrane protein with so-far-unknown function increased the dodecanoic acid methyl ester oxygenation activity of recombinantE. coli28-fold. In a two-liquid-phase bioreactor setup, a 62-fold increase to a maximal activity of 87 U gcdw−1was achieved, enabling the accumulation of high titers of terminally oxyfunctionalized products. Coexpression ofalkLalso increased oxygenation activities toward the natural AlkBGT substrates octane and nonane, showing for the first time clear evidence for a prominent role of AlkL in alkane degradation. This study demonstrates that AlkL is an efficient tool to boost productivities of whole-cell biotransformations involving hydrophobic aliphatic substrates and thus has potential for broad applicability.

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