The Journal of General and Applied Microbiology
Online ISSN : 1349-8037
Print ISSN : 0022-1260
ISSN-L : 0022-1260
Advance online publication
Displaying 1-18 of 18 articles from this issue
  • Ayako Kuri, Hirokazu Iida, Kazuyoshi Kawahara
    Article type: research-article
    Article ID: 2022.12.001
    Published: 2023
    Advance online publication: December 08, 2022
    JOURNAL FREE ACCESS ADVANCE PUBLICATION

    Cellular lipids of Lactiplantibacillus species were extracted and neutral glyceroglycolipids (GGLs) were purified, and analyzed by thin-layer chromatography (TLC). Four GGLs with known structures were detected in GGL preparation of L. plantarum, and the same GGL profiles of TLC were observed for all other strains of Lactiplantibacillus species examined, suggesting that the GGL profile could be one of the chemotaxonomic characters of the genus Lactiplantibacillus. On the other hand, the quantity of each GGL showed some variation among species, and L. pentosus was found to have higher proportion of disaccharide-type GGL, designated GGL-3 in this study, compared with other species including L. plantarum. The quantitative difference of GGL-3 found in this study could be regarded as the characteristics of L. pentosus. The carbohydrate structure of L. pentosus GGL-3 was precisely analyzed by 1H NMR and methylation analysis, and the structure was confirmed to be αGal-(1→2)-αGlc-diacylglycerol, with the carbohydrate structure identical to that of L. plantarum, although fatty acid composition of the two GGL-3 showed some difference.

    Download PDF (4544K)
  • Kazuki Ohta, Tenma Shimizu, Taku Oshima, Norikazu Ichihashi
    Article type: research-article
    Article ID: 2022.10.003
    Published: 2022
    Advance online publication: November 15, 2022
    JOURNAL FREE ACCESS ADVANCE PUBLICATION
    Supplementary material

    Various bacteria can change to a spherical cell-wall-deficient state, called L-from, in the presence of antibiotics that inhibit cell wall synthesis. L-forms are classified into two types: unstable and stable L-forms. Unstable L-forms revert to a normal walled state in the absence of antibiotics, while stable L-forms remain in their wall-deficient state. The conversion from unstable to stable L-forms has been often observed during long-term cultivation. However, the genetic cause for this conversion is not yet fully understood. Here, we obtained stable Bacillus subtilis L-form strains from unstable L-form strains via three independent long-term culturing experiments. The whole genome sequencing of the long-cultured strains identified many mutations, and some mutations were commonly found in all three long-cultured strains. The knockout strain of one of the commonly mutated genes, tagF, in the ancestral strain lost the ability to revert to walled state (rod shape), supporting that eliminating the function of tagF gene is one of the possible methods to convert unstable L forms to a stable state.

    Download PDF (624K)
  • Yuki Umebayashi, Shiori Abe, Miwa Yamada
    Article type: research-article
    Article ID: 2022.10.004
    Published: 2022
    Advance online publication: November 15, 2022
    JOURNAL FREE ACCESS ADVANCE PUBLICATION

    Cobetia bacteria are considered useful hosts for industrial applications owing to their fast growth, high cell density, and halophilicity. Here, we constructed an efficient conjugation method to obtain Cobetia sp. IU180733JP01 (5-11-6-3) transformants, which can produce bioplastics from alginate or seaweed waste. Lysogeny Broth medium containing 2% NaCl was used to co-cultivate the 5-11-6-3 strain (plasmid recipient) with Escherichia coli S17-1 (plasmid donor). Transformants with the highest conjugation efficiency [(2.92 ± 1.37) × 10-3] were obtained at a donor:recipient cell number ratio of 5:1. This is the first study reporting the creation of recombinant strains in the genus Cobetia. This method will contribute to creating a platform strain for the production of bioplastics and other useful materials from marine biomass.

    Download PDF (593K)
  • William Kurdy, Galina Yakovleva, Olga Ilinskaya
    Article type: research-article
    Article ID: 2022.10.001
    Published: 2022
    Advance online publication: November 02, 2022
    JOURNAL FREE ACCESS ADVANCE PUBLICATION
    Supplementary material

    Arctic ecosystems are affected by negative influence of climate change, pollution, and overexploitation of resources. Microorganisms playing a key role in preserving extreme econiches are poorly studied and require the use of modern methods for studying both their biodiversity and physiological activity. We applied Illumina MiSeq to the high-throughput 16S rRNA sequencing study of four Laptev Sea sediments from 64 - 185 m depth, using next generation sequencing enables rapid analysis of composition and diversity of prokaryotic communities. Although the dominant phylum in all samples was Proteobacteria, only the deepest sample contained a high number of archaeal organisms (19%) with the predominance of Methanosarcinaceace family in comparison with less 1% in the other three samples. This deepest sample had the lowest biodiversity and richness indices. Comparison of functional profiles of communities using Global Mapper tool revealed similar average abundance of infectiousness, drug resistance and environmental adaptation determinants in all samples, and high functional abundance for xenobiotic degradation in two samples. Among cultivated bacteria which could be promising producers of secreted RNase the representatives of Bacillus and Lysinibacillus genera were found. Our results contribute to improve our understanding of richness and ecological role of Laptev Sea microbiota.

    Download PDF (718K)
  • Yoshinori Tsuji, Akari Kinoshita, Mizuho Tsukahara, Takumi Ishikawa, H ...
    Article type: research-article
    Article ID: 2022.08.001
    Published: 2022
    Advance online publication: August 24, 2022
    JOURNAL FREE ACCESS ADVANCE PUBLICATION
    Supplementary material

    Yet another kinase (YAK) 1 is a conserved eukaryotic protein kinase coordinating growth and development. We previously isolated a mutant of Chlamydomonas reinhardtii defective in the YAK1 ortholog triacylglycerol (TAG) accumulation regulator 1 (TAR1). The mutant tar1-1 displayed higher levels of chlorophyll, starch, TAG, and biomass than the parental strain C9 (renamed as C9-3) in photoautotrophic nitrogen (N)-deficient conditions. However, we found that the parental C9-3 showed faster chlorosis upon N-deficiency than the original C9 (C9-1) freshly recovered from cryopreservation, suggesting that C9-3 had acquired particular characteristics during long-term subculturing. To exclude phenotypes dependent on a particular parental strain, we newly created tar1 mutants from two wild-types, C9-1 and CC 125. Like tar1-1, the new tar1 mutants showed higher levels of chlorophyll and TAG/starch than the parental strain. Upon removal of N, Chlamydomonas cells divide once before ceasing further division. Previously, the single division after N-removal was arrested in tar1-1 in photomixotrophic conditions, but this phenotype was not observed in photoautotrophic conditions because of the particular characteristics of the parental C9-3. However, using C9- 1 and CC-125 as parental strains, we showed that cell division after N-removal was impaired in new tar1 mutants in photoautotrophic conditions. Consistent with the view that the division under N-deficiency is necessary for gametic differentiation, new tar1 mutants showed lower mating efficiency than the parental strains. Taken together, TAR1 was suggested to promote differentiation into gametes through the regulation of cell division in response to N-deficiency.

    Download PDF (2519K)
  • Naoya Kataoka, Mienosuke Matsutani, Kazunobu Matsushita, Toshiharu Y ...
    Article type: research-article
    Article ID: 2022.08.002
    Published: 2022
    Advance online publication: August 22, 2022
    JOURNAL FREE ACCESS ADVANCE PUBLICATION
    Supplementary material

    Corynebacterium glutamicum was metabolically engineered to produce phenylalanine, a valuable aromatic amino acid that can be used as a raw material in the food and pharmaceutical industries. First, a starting phenylalanine-producer was constructed by overexpressing tryptophan-sensitive 3-deoxy-D-arabino-heptulosonate-7-phosphate synthase and phenylalanine- and tyrosine-insensitive bifunctional enzyme chorismate mutase prephenate dehydratase from Escherichia coli, followed by the inactivation of enzymes responsible for the formation of dihydroxyacetone and the consumption of shikimate pathway-related compounds. Second, redirection of the carbon flow from tyrosine to phenylalanine was attempted by deleting of the tyrA gene encoding prephenate dehydrogenase, which catalyzes the committed step for tyrosine biosynthesis from prephenate. However, suppressor mutants were generated, and two mutants were isolated and examined for phenylalanine production and genome sequencing. The suppressor mutant harboring an amino acid exchange (L180R) on RNase J, which was experimentally proven to lead to a loss of function of the enzyme, showed significantly enhanced production of phenylalanine. Finally, modifications of phosphoenolpyruvate-pyruvate metabolism were investigated, revealing that the inactivation of either phosphoenolpyruvate carboxylase or pyruvate carboxylase, which are enzymes of the anaplerotic pathway, is an effective means for improving phenylalanine production. The resultant strain, harboring a phosphoenolpyruvate carboxylase deficiency, synthesized 50.7 mM phenylalanine from 444 mM glucose. These results not only provided new insights into the practical mutations in constructing a phenylalanine-producing C. glutamicum but also demonstrated the creation of a potential strain for the biosynthesis of phenylalanine-derived compounds represented by plant secondary metabolites.

    Download PDF (541K)
  • Tomoko Fujiwara, Atsuko Matsura, Momoka Fukuda, Katsuaki Kuroki, Tomoo ...
    Article type: research-article
    Article ID: 2022.07.002
    Published: 2022
    Advance online publication: August 08, 2022
    JOURNAL FREE ACCESS ADVANCE PUBLICATION
    Supplementary material

    There is currently great interest in the salt-tolerant yeast strains used to produce miso and soy sauce. Since the isolation of Zygosaccharomyces sp. strain from Japanese miso more than 60 years, several hybrid strains have been identified in fermented foods. Studies have shown that the active mating-type locus of the original Zygosaccharomyces sp. yeast strain is located between the T-subgenome sequence and the P-subgenome sequence. In this study, 32 salt-tolerant Zygosaccharomyces sp. yeast strains were isolated from five miso factories in Hiroshima Prefecture, Japan. Analysis by flow cytometry revealed that 27 strains were diploid and five strains were haploid. PCR analysis indicated that the 27 diploid strains had the same chromosomal structure of the active mating-type (MAT) locus as the original yeast strain isolated from miso 60 years ago. In addition, the 27 diploid strains were allodiploid, namely, natural hybrids of Z. rouxii and a related species, while the five haploid strains were all Z. rouxii. We found that cells of yeast strains isolated from miso changed morphologically when co-cultured with a yeast strain of opposite mating-type under nitrogen starvation conditions. The DNA sequence of the active mating-type locus and the results of cell morphology changes by co-culture were consistent with the mating type of each strain shown in the mating experiments. These findings will be useful for the future production of miso and soy sauce.

    Download PDF (3104K)
  • Sarunpron Khruengsai, Teerapong Sripahco, Patcharee Pripdeevech
    Article type: research-article
    Article ID: 2022.07.001
    Published: 2022
    Advance online publication: August 04, 2022
    JOURNAL FREE ACCESS ADVANCE PUBLICATION

    Low-density polyethylene (LDPE) has been commercially used and accumulated as plastic solid waste. LDPE has also been found to be a non-degradable waste for decades and found as a pollution source in the environment. In this study, 65 fungi were screened for their biodegradation of LDPE. The fungi Neopestalotiopsis phangngaensis, Alternaria burnsii, Alternaria pseudoeichhorniae, and Arthrinium sacchari showed significant potential in LDPE biodegradation. These fungi were individually cultured with an LDPE sheet as a carbon source for 90 days. A maximum weight loss of the LDPE sheet was detected by the fungus N. phangngaensis (54.34%). This fungus also revealed the highest reduction rate of tensile strength of the LDPE sheet (0.33 MPa). The morphological surface of LDPE culturing with N. phangngaensis was crack, pit, and rough analyzed by scanning electron microscopy. The biodegradation of the LDPE sheet by N. phangngaensis was also confirmed by the Sturm test and analysis of enzymatic activities. The Sturm test showed the highest decomposition of the LDPE sheet by N. phangngaensis into CO2 with 2.14 g/L after incubation. Enzymatic activities of laccase, manganese peroxidase, and lignin peroxidase enzymes were found by N. phangngaensis during the LDPE degradation. The volatile organic compounds in culture supernatant of N. phangngaensis were also investigated. The major compounds were 3Z-diethyl acetal hexenal, 2E,4E-decadienol, and 2Z-diethyl acetal hexenal. This study reveals the utilization of the fungus N. phangngaensis as the carbon source at a considerable biodegradation rate without any prior treatment. Therefore, the fungus N. phangngaensis may be applied as an alternative degrader for LDPE degradation in the environment.

    Download PDF (539K)
  • Suma Devivilla, Manjusha Lekshmi, Fathima Salam, Sanath Kumar H, Ra ...
    Article type: research-article
    Article ID: 2022.04.001
    Published: 2022
    Advance online publication: July 21, 2022
    JOURNAL FREE ACCESS ADVANCE PUBLICATION

    Consumption of temperature-abused marine fish containing elevated levels of histamine results in histamine poisoning. Histamine is a biogenic amine produced in fish by the action of certain groups of bacteria which are capable of producing an exogenous enzyme called histidine decarboxylase (HDC). Morganella morganii is one of the major causative organisms of histamine poisoning. In this study, the histamine forming potential of M. morganii (BSS142) is evaluated when it is co-incubated with proteolytic and polyamine forming bacteria. This experiment was designed to examine whether biotic factors such as proteolysis and the presence of other amines influenced histamine forming ability of BSS142. The study showed that the proteolytic activity of Aeromonas hydrophila as well as Pseudomonas aeruginosa greatly enhanced the histamine forming ability of M. morganii. Psychrobacter sangunis, a non proteolytic polyamine producer, negatively influenced histamine production by M. morganii.

    Download PDF (430K)
  • Ruijie Wang, Manabu Arioka
    Article type: research-article
    Article ID: 2022.06.002
    Published: 2022
    Advance online publication: July 21, 2022
    JOURNAL FREE ACCESS ADVANCE PUBLICATION
    Supplementary material

    Glucuronoyl esterase (GE) is a promising agent for the delignification of plant biomass since it has been shown to cleave the linkage between xylan and lignin in vitro. In this study, we demonstrate that NcGE, a GE from Neurospora crassa, stimulates plant biomass degradation. In vitro, NcGE synergistically increased the release of reducing sugars from plant biomass when added together with cellulase or xylanase. In vivo, overexpression of NcGE in N. crassa resulted in an increase in xylanolytic activity. Consistently, elevated transcription of genes encoding the major plant biomass degrading-enzymes (PBDEs) was observed in the NcGE overexpression strain. Increased xylanolytic activity and transcription of PDBE genes were largely abolished when the transcription factors clr-1, clr-2, or xlr-1 were deleted. Interestingly, the expression of some PBDE genes was increased when the hydrolysate of plant biomass by NcGE was added to the culture medium. We propose that NcGE boosts the production of PBDEs through the activation of key transcription factors, which is presumably caused by NcGE-mediated generation of hypothetical inducer(s) from plant biomass.

    Download PDF (601K)
  • Qiong Deng, Zhu Wang, Pengmei Wu, Hui Liang, Haixia Wu, Lirong Zhang, ...
    Article type: research-article
    Article ID: 2022.05.004
    Published: 2022
    Advance online publication: July 09, 2022
    JOURNAL FREE ACCESS ADVANCE PUBLICATION
    Supplementary material

    Mycoplasma pneumoniae is one of the most important pathogens causing community acquired pneumonia in children, and the pathogenic mechanism of M. pneumoniae infection is complex. Azithromycin is an effective agent for treating the acquired lower respiratory tract infection and urogenital tract infection with slight adverse reactions. This study aimed to compare the intestinal microflora before (PP1) and after azithromycin intervention (PP2) in children with pneumonia caused by M. pneumoniae, combined with body fluid biochemical analysis to determine the intestinal flora affecting the progress of the disease. Fifteen children diagnosed with M. pneumoniae pneumonia were recruited. The fecal samples and clinical biochemical data were collected. 16S rRNA gene amplicon sequencing and bioinformatics analysis were conducted by the Beijing Genomics Institute. The operational taxonomic unit abundance analysis showed significant differences between the two groups. The species richness analysis showed differences in class, family, genus, order, species, and phylum. The abundance of Haemophilus, Pasteurellales, and Pasteurellaceae was found to be significantly higher in the PP1 group. The Pearson correlation analysis showed that the microbes strongly correlated with the clinical features. 16S rRNA gene sequencing data revealed altered composition of gut microbiota in children with M. pneumoniae pneumonia treated with azithromycin. The altered expression of microbes correlated with clinical features, which might help diagnose and treat the disease.

    Download PDF (2110K)
  • Tomohiro Inaba, Mami Yamaguchi, Akira Taniguchi, Yuya Sato, Tomo Aoy ...
    Article type: research-article
    Article ID: 2022.04.003
    Published: 2022
    Advance online publication: June 13, 2022
    JOURNAL FREE ACCESS ADVANCE PUBLICATION
    Supplementary material

    The decolorization of 11 dyes by granular sludge from an anaerobic expanded granular sludge bed (EGSB) reactor was evaluated. Biological decolorization of Reactive Red 21, 23, and 180, and Reactive Yellow 15, 17, and 23 in model textile wastewater was observed for the first time after a 7-day incubation (over 94% decolorization). According to the sequencing analysis of 16S rRNA gene amplicons from EGSB granular sludge, the operational taxonomic unit related to Paludibacter propionicigenes showed the highest increase in relative abundance ratios in the presence of dyes (7.12 times on average over 11 dyes) compared to those without dyes.

    Download PDF (775K)
  • Vu Dinh Giap, Hoang Thanh Duc, Pham Thi Mai Huong, Do Thi Hanh, Do H ...
    Article type: research-article
    Article ID: 2022.05.005
    Published: 2022
    Advance online publication: July 02, 2022
    JOURNAL FREE ACCESS ADVANCE PUBLICATION

    From the biotechnological point of view, enzymes are powerful tools that help sustain a clean environment in several ways. The enzymatic biodegradation of synthetic dyes is a promising goal since it reduces pollution caused by textile dyeing factory wastewater. Lignin peroxidase (EC 1.11.1.14, LiP) has high redox potential; thus, it is great for application in various industrial fields (e.g., paper- waste treatment and textile dyeing wastewater treatment). In the present study, a LiP from an isolated strain Pleurotus pulmonarius CPG6 (PpuLiP) was successfully purified with a specific activity of 6.59 U mg -1. The enzyme was purified by using three-step column chromatography procedures including DEAE, Sephadex G-75, and HiTrapTM Q FF columns with 17.8-fold purity. The enzyme with a molecular weight of 40 kDa exhibited enhanced pH stability in the acidic range. The activity retention was over 75% at a pH of 3.0 for more than 6 hours. Purified PpuLiP was able to oxidize a variety of substrates including veratryl alcohol, 2,4-DCP, n propanol, and guaiacol. The effect of metal ions on PpuLiP activity was analyzed. The study will provide a ground to decolorize dyes from various groups of PpuLiP. Purified PpuLiP could decolorize 35% Acid blue 25 (AB25), 50% Acid red 129 (AB129), 72% Acid blue 62 (NY3), 85% Acid blue 113 (AB113), 55% Remazol Brilliant blue R (RBBR), and 100% Reactive red 120 (RR120) for 12 hours. Most of the dyes were decolorized, but the heat-denatured enzyme used as negative control obviously did not decolorize the tested dyes. These results indicate that the PpuLiP has potential application in enzyme-based decolorization of synthetic dyes. Keywords: Decolorization; lignin peroxidase; Pleurotus pulmonarius; textile dyes.

    Download PDF (626K)
  • Teppei Maruyama, Kanako Hayashi, Kotaro Matsui, Yasukichi Maekawa, Tak ...
    Article type: research-article
    Article ID: 2022.05.006
    Published: 2022
    Advance online publication: July 02, 2022
    JOURNAL FREE ACCESS ADVANCE PUBLICATION
    Supplementary material

    Fission yeast, Schizosaccharomyces pombe, possesses eight hexose transporters, Ght1~8. In order to clarify the role of each hexose transporter on glucose uptake, a glucose uptake assay system was established and the actual glucose uptake activity of each hexose transporter-deletion mutant was measured. Under normal growth condition containing 2% glucose, ∆ght5 and ∆ght2 mutants showed large and small decrease in glucose uptake activity, respectively. On the other hand, the other deletion mutants did not show any decrease in glucose uptake activity indicating that, in the presence of Ght5 and Ght2, the other hexose transporters do not play a significant role in glucose uptake. To understand the relevance between glucose uptake and lifespan regulation, we measured the chronological lifespan of each hexose transporter deletion mutant, and found that only ∆ght5 mutant showed a significant lifespan extension. Based on these results we showed that Ght5 is mainly involved in the glucose uptake in Schizosaccharomyces pombe, and suggested that the ∆ght5 mutant has prolonged lifespan due to physiological changes similar to calorie restriction.

    Download PDF (1411K)
  • Yanxiang Yao, Naren Xi, E Hai, Xiaomin Zhang, Jiayi Guo, Zhi Lin, Wei ...
    Article type: research-article
    Article ID: 2022.05.003
    Published: 2022
    Advance online publication: June 21, 2022
    JOURNAL FREE ACCESS ADVANCE PUBLICATION
    Supplementary material

    As a central signaling molecule, c-di-GMP (bis-(3,5)-cyclic diguanosine monophosphate) is becoming the focus for research in bacteria physiology. Pseudomonas aeruginosa PAO1 genome contains highly complicated c-di-GMP metabolizing genes and a number of these proteins have been identified and investigated. Especially, a sophisticated network of these proteins is emerging. In current study, mainly through Bacteria-2-Hybrid assay, we found PA0575 (RmcA), a GGDEF-EAL dual protein, to interact with two other dual proteins of PA4601 (MorA) and PA4959 (FimX). These observations imply the intricacy of c-di-GMP metabolizing protein interactions. Our work thus provides one piece of data to increase the understandings to c-di-GMP signaling.

    Download PDF (1500K)
  • Orn anong Chaiyachet, Ketsara Wongtham, Komsan Sangkasame
    Article type: research-article
    Article ID: 2022.05.002
    Published: 2022
    Advance online publication: June 10, 2022
    JOURNAL FREE ACCESS ADVANCE PUBLICATION

    The present study investigated the efficacy of bacterial cellulose production by K. xylinus TISTR 1011 and K. nataicola TISTR 975 using yam bean juice as a nutrient source, and the physicochemical and sensory characteristics of bacterial cellulose were examined. Bacterial cellulose content, production yield, and production rate were significantly higher when K. xylinus TISTR 1011 rather than K. nataicola TISTR 975 was used as the bacterial strain. The analysis of physicochemical characteristics revealed that bacterial cellulose produced by K. xylinus TISTR 1011 using yam bean juice medium had higher scores for CIE L*, a*, and b* values, wet weight, moisture content, firmness, and gel strength than bacterial cellulose produced by K. nataicola TISTR 975. In contrast, sensory evaluation showed that the acceptability scores and preference of all attributes of bacterial cellulose produced by K. nataicola TISTR 975 using yam bean juice medium were higher than those of bacterial cellulose produced by K. xylinus TISTR 1011. The results of this study indicate that yam bean juice from yam bean tubers, an alternative raw material agricultural product, can be used as a nutrient source for producing bacterial cellulose or nata by Komagataeibacter strains.

    Download PDF (353K)
  • Kasumi Shimodate, Hiroyuki Honda
    Article type: research-article
    Article ID: 2022.04.002
    Published: 2022
    Advance online publication: June 08, 2022
    JOURNAL FREE ACCESS ADVANCE PUBLICATION

    Godo is a traditional fermented soy food made in Aomori prefecture, Japan. It is mainly made of soybeans, rice koji, and salt. Since godo ripens during the long and severe winter in northeast Japan, it is assumed that lactic acid bacteria inhabiting godo have cold tolerance. We aimed to investigate the presence or absence of psychrotrophic lactic acid bacteria in godo. The viable counts of estimated lactic acid bacteria ranged from 106 to 108 cfu/g. In addition, aerobic and anaerobic microorganisms were detected in four godo products though the microbial population differed from sample to sample. Twenty-two bacterial strains were able to be isolated from godo, and all of the isolated strains were Gram-positive and catalase-negative. Some of the isolates grew well at 10°C. The carbohydrate fermentation profile of the selected three strains was determined by API50 CHL analysis. These strains were identified as Leuconostoc mesenteroides, and Latilactobacillus sakei by 16S rRNA gene sequence analysis. Leuconostoc mesenteroides strains HIT231 and HIT252, and Latilactobacillus sakei strain HIT273 could grow at 5°C in MRS broth, but their optimum growth temperature was 20°C-30°C. These results suggest that psychrotrophic lactic acid bacteria presumed to be derived from rice koji are present in godo, which is one of the factors in the low temperature ripening of godo in winter.

    Download PDF (1609K)
  • Takashi Kuribayashi, Toshiki Sakurai, Akira Hatakeyama, Toshio Joh, Mi ...
    Article type: research-article
    Article ID: 2022.05.001
    Published: 2022
    Advance online publication: June 08, 2022
    JOURNAL FREE ACCESS ADVANCE PUBLICATION

    In Saccharomyces cerevisiae, ethyl caprylate is produced by the esterification of caprylic acid, which is synthesized through the action of fatty acid synthase. A recent study reported a yeast mutant with a single nucleotide substitution in the alpha subunit of fatty acid synthase (FAS2) gene (F1279Y; 3836T>A) that produced large amounts of ethyl caprylate. Here, we designed two primer sets (P1/P2 and P3/P4) with mismatches that incorporate restriction sites for the enzymes NdeI and SspI, respectively and developed an easy and rapid polymerase chain reaction-restriction fragment length polymorphism assay to identify yeasts harboring the FAS2-F1279Y mutation associated with high ethyl caprylate productivity.

    Download PDF (659K)
feedback
Top