Engineering Escherichia coli BL21 genome to improve the heptanoic acid tolerance by using CRISPR-Cas9 system
Acid tolerance is one of the critical factors to determine the quality of the industrial production strains. Therefore, we have investigated the introduction of the acid tolerance genes into the genome of Escherichia coli BL21 by using CRISPR-Cas9 system. The dsrA and rcsB genes of E. coli K-12, whi...
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| Published in | Biotechnology and bioprocess engineering Vol. 22; no. 3; pp. 231 - 238 |
|---|---|
| Main Authors | , , , |
| Format | Journal Article |
| Language | English |
| Published |
Seoul
The Korean Society for Biotechnology and Bioengineering
01.06.2017
Springer Nature B.V 한국생물공학회 |
| Subjects | |
| Online Access | Get full text |
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| Abstract | Acid tolerance is one of the critical factors to determine the quality of the industrial production strains. Therefore, we have investigated the introduction of the acid tolerance genes into the genome of
Escherichia coli
BL21 by using CRISPR-Cas9 system. The
dsrA
and
rcsB
genes of
E. coli
K-12, which are involved in the heptanoic acid tolerance, were inserted into the genome of
E. coli
BL21 without scar. The native transcription unit (TU) of dsrA and the synthetic TU of
rcsB
were integrated in E. coli BL21 genome. We found that the position of genomic coordinate of 1,300,270 was more efficient to integrate dsrA and
rcsB
than genomic coordinate of 3,876,428. Furthermore, the
rcsB
was successfully expressed in the resulting engineered strains (
i.e
.,
rcsB
+
or
dsrA
+
rcsB
+
strains). The engineered strains expressing
dsrA
and/or
rcsB
showed the higher survival rate and specific growth rate under
n
-heptanoic acid stress than wild-type
E. coli
BL21. These results indicate that the newly introduced acid-tolerance systems were active in the
E. coli
BL21 strain. |
|---|---|
| AbstractList | Acid tolerance is one of the critical factors to determine the quality of the industrial production strains. Therefore, we have investigated the introduction of the acid tolerance genes into the genome of
Escherichia coli
BL21 by using CRISPR-Cas9 system. The
dsrA
and
rcsB
genes of
E. coli
K-12, which are involved in the heptanoic acid tolerance, were inserted into the genome of
E. coli
BL21 without scar. The native transcription unit (TU) of dsrA and the synthetic TU of
rcsB
were integrated in E. coli BL21 genome. We found that the position of genomic coordinate of 1,300,270 was more efficient to integrate dsrA and
rcsB
than genomic coordinate of 3,876,428. Furthermore, the
rcsB
was successfully expressed in the resulting engineered strains (
i.e
.,
rcsB
+
or
dsrA
+
rcsB
+
strains). The engineered strains expressing
dsrA
and/or
rcsB
showed the higher survival rate and specific growth rate under
n
-heptanoic acid stress than wild-type
E. coli
BL21. These results indicate that the newly introduced acid-tolerance systems were active in the
E. coli
BL21 strain. Acid tolerance is one of the critical factors to determine the quality of the industrial production strains. Therefore, we have investigated the introduction of the acid tolerance genes into the genome of Escherichia coli BL21 by using CRISPR-Cas9 system. The dsrA and rcsB genes of E. coli K-12, which are involved in the heptanoic acid tolerance, were inserted into the genome of E. coli BL21 without scar. The native transcription unit (TU) of dsrA and the synthetic TU of rcsB were integrated in E. coli BL21 genome. We found that the position of genomic coordinate of 1,300,270 was more efficient to integrate dsrA and rcsB than genomic coordinate of 3,876,428. Furthermore, the rcsB was successfully expressed in the resulting engineered strains (i.e., rcsB+ or dsrA+ rcsB+ strains). The engineered strains expressing dsrA and/or rcsB showed the higher survival rate and specific growth rate under n-heptanoic acid stress than wild-type E. coli BL21. These results indicate that the newly introduced acid-tolerance systems were active in the E. coli BL21 strain. KCI Citation Count: 8 Acid tolerance is one of the critical factors to determine the quality of the industrial production strains. Therefore, we have investigated the introduction of the acid tolerance genes into the genome of Escherichia coli BL21 by using CRISPR-Cas9 system. The dsrA and rcsB genes of E. coli K-12, which are involved in the heptanoic acid tolerance, were inserted into the genome of E. coli BL21 without scar. The native transcription unit (TU) of dsrA and the synthetic TU of rcsB were integrated in E. coli BL21 genome. We found that the position of genomic coordinate of 1,300,270 was more efficient to integrate dsrA and rcsB than genomic coordinate of 3,876,428. Furthermore, the rcsB was successfully expressed in the resulting engineered strains (i.e., rcsB + or dsrA + rcsB + strains). The engineered strains expressing dsrA and/or rcsB showed the higher survival rate and specific growth rate under n-heptanoic acid stress than wild-type E. coli BL21. These results indicate that the newly introduced acid-tolerance systems were active in the E. coli BL21 strain. |
| Author | Baek, So-Won Lee, Jinwon Park, Jin-Byung Seo, Joo-Hyun |
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| CitedBy_id | crossref_primary_10_1007_s12257_018_0374_6 crossref_primary_10_1007_s12257_018_0379_1 crossref_primary_10_1007_s12257_018_0252_2 crossref_primary_10_1016_j_jbiotec_2018_07_019 crossref_primary_10_1002_biot_201700596 crossref_primary_10_1002_bit_28404 crossref_primary_10_1016_j_engmic_2023_100101 crossref_primary_10_1021_acssynbio_8b00519 crossref_primary_10_1186_s12934_022_01746_z crossref_primary_10_1016_j_bej_2021_108254 crossref_primary_10_3390_microorganisms11040907 crossref_primary_10_1039_C9CY01802F crossref_primary_10_1016_j_indcrop_2024_118937 crossref_primary_10_1128_mbio_03656_21 crossref_primary_10_1007_s00253_018_9503_6 crossref_primary_10_1016_j_mimet_2022_106648 |
| Cites_doi | 10.1038/nbt.3365 10.1016/j.molcel.2009.06.035 10.1371/journal.pone.0163265 10.1016/j.ymben.2015.06.006 10.1261/rna.5183804 10.1007/s12257-015-0607-x 10.3389/fmicb.2013.00272 10.1002/adsc.201600216 10.1007/s00253-009-2355-3 10.1101/gr.162339.113 10.1038/srep34651 10.1021/acscatal.6b01884 10.1093/nar/gkt651 10.1038/srep28223 10.1039/C5GC01017A 10.1021/acssynbio.5b00187 10.1038/ncomms14665 10.1016/S0958-1669(05)80142-9 10.1128/JB.05040-11 10.1128/JB.186.18.6179-6185.2004 10.1093/nar/gkq097 10.1128/AEM.04023-14 10.1093/nar/gku828 |
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| Title | Engineering Escherichia coli BL21 genome to improve the heptanoic acid tolerance by using CRISPR-Cas9 system |
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