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The Prokaryotic Microalga Limnothrix redekei KNUA012 to Improve Aldehyde Decarbonylase Expression for Use as a Biological Resource Cover

The Prokaryotic Microalga Limnothrix redekei KNUA012 to Improve Aldehyde Decarbonylase Expression for Use as a Biological Resource

Polish Journal of Microbiology
Volume 72 (2023): Issue 3 (September 2023)
By:
Open Access
|Sep 2023

Figures & Tables

Fig. 1.

Phylogenetic relationships of Limnothrix redekei KNUA012 and related organisms.The tree is constructed to scale with branch lengths in the same units as those of the evolutionary distances used to infer the phylogenetic tree. The evolutionary distances were computed using the maximum composite likelihood method (Tamura et al. 2004), and evolutionary analyses were conducted using MEGA5 (Tamura et al. 2011).
Phylogenetic relationships of Limnothrix redekei KNUA012 and related organisms.The tree is constructed to scale with branch lengths in the same units as those of the evolutionary distances used to infer the phylogenetic tree. The evolutionary distances were computed using the maximum composite likelihood method (Tamura et al. 2004), and evolutionary analyses were conducted using MEGA5 (Tamura et al. 2011).

Fig. 2.

a) Light microscopy images of Limnothrix redekei KNUA012. The cell diameter ranged from 10 μm depending on the growth stage. Live cells were visualized at magnifications of 400 × (left panel) and 1,000 × (right panel) under a microscope equipped with differential interference contrast optics.b) Growth curves of Limnothrix redekei KNUA012 at 5–35°C (at intervals of 5°C).
a) Light microscopy images of Limnothrix redekei KNUA012. The cell diameter ranged from 10 μm depending on the growth stage. Live cells were visualized at magnifications of 400 × (left panel) and 1,000 × (right panel) under a microscope equipped with differential interference contrast optics.b) Growth curves of Limnothrix redekei KNUA012 at 5–35°C (at intervals of 5°C).

Fig. 3.

SDS polyacrylamide gel images of the aldehyde decarbonylase (AD) protein purified from Escherichia coli cells transformed with the expression vector pET-28a carrying AD.a) M – marker, lane 1 – protein purified from E. coli cells transformed with pET-28a (pET28 empty vector), lane 2 – protein purified from E. coli cells transformed with pET28-AD (pET-28a harboring the AD gene); b) M – marker, lane 3 – purified AD obtained from E. coli (pET28-AD).
SDS polyacrylamide gel images of the aldehyde decarbonylase (AD) protein purified from Escherichia coli cells transformed with the expression vector pET-28a carrying AD.a) M – marker, lane 1 – protein purified from E. coli cells transformed with pET-28a (pET28 empty vector), lane 2 – protein purified from E. coli cells transformed with pET28-AD (pET-28a harboring the AD gene); b) M – marker, lane 3 – purified AD obtained from E. coli (pET28-AD).

Fig. 4.

Analysis of the GC peaks.a) GC/MS total ion chromatogram of standard fatty acid methyl esters. 1 – Tridecanoic acid methyl ester, 2 – pentadecanoic acid methyl ester, 3 – heptadecanoic acid methyl ester, 4 – nonadecanoic acid methyl ester, and 5 – henelcosanoic acid methyl ester.b) GC peak results for the fatty acids extracted from L. redekei KNUA012. 1 – Pentadecane, 2 – dodecanoic acid methyl ester, 3 – 8-heptadecene, 4 – heptadecane, 5 – methyl Z-11-tetradecenoate, 6 – tetradecanoic acid methyl ester, 7 – 9-hexadecenoic acid methyl ester, 8 – palmitoleic acid methyl ester, 9 – hexadecanoic acid methyl ester, 10 – 9-octadecenoic acid methyl ester, and 11 – octadecanoic acid methyl ester. Peak no. annotated in Table SI
Analysis of the GC peaks.a) GC/MS total ion chromatogram of standard fatty acid methyl esters. 1 – Tridecanoic acid methyl ester, 2 – pentadecanoic acid methyl ester, 3 – heptadecanoic acid methyl ester, 4 – nonadecanoic acid methyl ester, and 5 – henelcosanoic acid methyl ester.b) GC peak results for the fatty acids extracted from L. redekei KNUA012. 1 – Pentadecane, 2 – dodecanoic acid methyl ester, 3 – 8-heptadecene, 4 – heptadecane, 5 – methyl Z-11-tetradecenoate, 6 – tetradecanoic acid methyl ester, 7 – 9-hexadecenoic acid methyl ester, 8 – palmitoleic acid methyl ester, 9 – hexadecanoic acid methyl ester, 10 – 9-octadecenoic acid methyl ester, and 11 – octadecanoic acid methyl ester. Peak no. annotated in Table SI

GC/MS results showing the alkanes and major fatty acids present in Limnothrix redekei KNUA012, Escherichia coli cells transformed with pET28-AD, and the empty vector pET28_

Peak no.Component namepET28 empty vector (% w/w)KNUA012 (% w/w)pET28-AD (% w/w)
1Pentadecane0.87 ± 0.522.40 ± 0.851.78 ± 0.38
2Dodecanoic acid methyl ester0.18 ± 0.050.54 ± 0.210.25 ± 0.08
38-Heptadecene0.54 ± 0.070.62 ± 0.231.09 ± 0.31
4Heptadecane1.07 ± 0.171.15 ± 0.421.86 ± 0.52
5Methyl Z-11-tetradecenoate21.7 ± 1.5222.1 ± 1.0323.6 ± 1.59
6Tetradecanoic acid methyl ester11.2 ± 1.3111.2 ± 1.0715.7 ± 1.41
79-Hexadecenoic acid methyl ester2.62 ± 0.8421.3 ± 1.518.63 ± 1.52
8Palmitoleic acid methyl ester2.35 ± 0.535.37 ± 1.033.82 ± 0.82
9Hexadecanoic acid methyl ester2.06 ± 0.5121.7 ± 1.857.82 ± 1.31
109-Octadecenoic acid methyl ester3.64 ± 0.859.31 ± 1.376.83 ± 1.46
11Octadecanoic acid methyl ester0.84 ± 0.061.04 ± 0.521.83 ± 0.82
DOI: https://doi.org/10.33073/pjm-2023-031 | Journal eISSN: 2544-4646 | Journal ISSN: 1733-1331
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Language: English
Page range: 307 - 317
Submitted on: Mar 15, 2023
Accepted on: Jul 4, 2023
Published on: Sep 20, 2023
Published by: Polish Society of Microbiologists
In partnership with: Paradigm Publishing Services
Publication frequency: 4 times per year
Keywords:
alkanes,
aldehyde decarbonylase,
biofuels,
biomass,
fatty acid
Related subjects:
Life sciences,
Microbiology and virology

© 2023 Young-Saeng Kim, Haeri Baek, Hyun-Sik Yun, Jae-Hak Lee, Kyoung-In Lee, Han-Soon Kim, Ho-Sung Yoon, published by Polish Society of Microbiologists
This work is licensed under the Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 License.

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