Isolation and Identification of Acetic Acid Bacteria for Submerged Acetic Acid Fermentation at High Temperature

Ohmori, Shouji; Masai, Hiroshi; Arima, Kei; Beppu, Teruhiko

doi:10.1080/00021369.1980.10864432

Cited by 21 publications

(23 citation statements)

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“…The concentration of acetic acid was measured by titration with 1 N NaOH as described by Ohmori et al (15 ability of each strain thus obtained was determined by measuring the production of acetic acid as described by Ohmori et al (16). The resistance to acetic acid was determined by the method described by Ohmori et al (15).…”

Section: Methodsmentioning

confidence: 99%

Novel insertion sequence IS1380 from Acetobacter pasteurianus is involved in loss of ethanol-oxidizing ability

1991

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Acetobacter pasteurianus NCI1380, a thermophilic strain isolated from the surface culture of acetic acid fermentation, showed genetic instability to produce at high frequency spontaneous mutants which were deficient in ethanol oxidation because of the loss of alcohol dehydrogenase activity. Southern hybridization experiments with the cloned alcohol dehydrogenase-cytochrome c gene cluster as the probe showed insertion of an unknown DNA fragment into a specific position in the cytochrome c gene in most of the mutant strains. Cloning and sequencing analyses revealed that the inserted sequence was 1,665 bp in length and had a terminal inverted repeat of 15 bp. In addition, this inserted sequence was found to generate a 4-bp duplication at the inserted site upon transposition. The target site specificity was not very strict, but a TCGA sequence appeared to be preferentially used. The inserted sequence contains two long open reading frames of 461 and 222 amino acids which are overlapped and encoded by different strands. Although these open reading frames showed no homology to any protein registered in the DNA data bases, the longer open reading frame contained many basic amino acids (87 of 461), as was observed with transposases of so-called insertion sequence (IS) elements. All of these characteristics are typical of IS elements, and the sequence was named IS1380. The copy number of IS1380 in a cell of A. pasteurianus NCI1380 was estimated to be about 100. Several strains of acetic acid bacteria also contained IS1380 at high copy numbers. These results suggest that IS1380 is associated with the genetic loss of ethanol-oxidizing ability as well as the genetic instability of acetic acid bacteria in general.

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Section: Methodsmentioning

confidence: 99%

Novel insertion sequence IS1380 from Acetobacter pasteurianus is involved in loss of ethanol-oxidizing ability

1991

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“…In addition to the genetic instability in ethanol oxidation in Acetobacter spp., spontaneous mutations resulting in deficiency in acetic acid resistance (Ohmori et al, 1980) and cellulose formation (Cook & Colvin, 1980;Steel & Walker, 1957) arise at high frequencies. Investigations have revealed that insertion sequences, such as IS1031s (from 1S1031A to IS1031D) and 19032, participate in the inactivation of several genes (Coucheron, 1991(Coucheron, , 1993Iversen et al, 1994).…”

Section: Introductionmentioning

confidence: 99%

A new insertion sequence IS1452 from Acetobacter pasteurianus

Kondo

Horinouchi

1997

Microbiology

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A new insertion sequence element, IS1452, was found to be associated with inactivation of the alcohol dehydrogenase by insertion in the adhS gene encoding subunit III of the three-component membrane-bound alcohol dehydrogenase complex in Acetobacter pasteurianus. Cloning and sequencing analyses of the mutated subunit III gene locus revealed that IS 1452 was inserted at or near the ribosome-binding sequence of adhS. Analysis of transcription using the chloramphenicol acetyltransferase gene as the reporter indicated that IS1452 abolished transcription of adhS by separating its promoter from the subunit III structural gene. IS1452 was 1411 bp in length and had a terminal inverted repeat of 21 bp. IS1452 contained one long ORF of 416 amino acids rich in basic amino acids. This protein showed homology with a putative transposase, Tra1, of IS701 isolated from the cyanobacterium Calothrix species PCC 7601. Like IS701, IS7452 was found to generate a 4 bp direct repeat at the site of insertion upon transposition. The target site specificity was rather strict, and a CTA(A or G) sequence appeared to be preferentially recognized. Transposition of IS1452 was replicative, since it was accompanied by an increase in the copy number of IS1452. Several strains belonging to the genus Acetobacter also contained IS1452 at varying copy numbers from one to more than ten. These observations suggest that IS1452 is one of the insertion sequences that are responsible for genetic instability leading to deficiencies in various physiological properties in acetic acid bacteria.

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“…To elucidate the genetic background of the terminal oxidase in ethanol oxidation, we first purified and characterized the terminal oxidase from a thermotolerant strain, A. aceti 1023 (24), a suitable strain for submerged acetic acid fermentation. We then cloned and characterized the gene cluster * Corresponding author.…”

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Characterization of a cytochrome a1 that functions as a ubiquinol oxidase in Acetobacter aceti

et al. 1993

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The terminal oxidase for ethanol oxidation in Acetobacter aced was purified as a complex consisting of four subunits (subunits I, II, III, and IV) with molecular masses of 72, 34, 21, and 13 kDa, respectively.Spectrophotometric analysis and catalytic properties determined with the purified enzyme showed that it belonged to a family of cytochrome a, (ba) (cyaB, cyaC, and cyaD) encoding the other three subunits (subunits II, III, and IV) were clustered upstream and downstream of the cyaA gene in the order cyaB, cyat4, cyaC, and cyaD and with the same transcription polarity, forming an operon. As expected from the enzymatic properties, CyaB, CyaC, and CyaD showed great similarity in amino acid sequence to the corresponding subunits of the E. coli o-type ubiquinol oxidase and aa3-tYpe cytochrome c oxidases.The characteristic activity of the genus Acetobacter in vinegar production is the oxidation of ethanol, which is catalyzed by the two membrane-associated dehydrogenases, alcohol dehydrogenase and aldehyde dehydrogenase. The enzymatic properties of these dehydrogenases have been elucidated (1, 2, 9, 34), and the genes encoding these enzymes have been cloned and sequenced (10,32,33). Electrons generated in ethanol oxidation are thought to be directly transferred through the respiratory chain to the terminal oxidase via ubiquinone (20). Acetobacter strains were divided into two groups on the basis of the terminal oxidases; one contains cytochrome a,, and the other contains cytochrome d (3). Williams and Poole (36, 37) observed that the amount of cytochrome o, as the terminal oxidase of Acetobacterpasteurianus NCIB 6428, increased under aerobic culture conditions. Recently, Matsushita et al. (17,19) reported that cytochrome al ofA. aceti, which is composed of four subunits (55,35,22, and 18 kDa) containing heme b, heme a, and one copper ion, functions as a ubiquinol oxidase. They also examined the effect of aeration on the terminal ubiquinol oxidase and observed that a change in culture conditions caused a change in the composition of the terminal ubiquinol oxidase (17, 18).To elucidate the genetic background of the terminal oxidase in ethanol oxidation, we first purified and characterized the terminal oxidase from a thermotolerant strain, A. aceti 1023 (24), a suitable strain for submerged acetic acid fermentation. We then cloned and characterized the gene cluster * Corresponding author.encoding all the subunits of the terminal oxidase. In this paper, we describe the enzymatic properties of the ubiquinol oxidase from A. aceti and the cloning and sequencing of the gene cluster. In addition, we compared the amino acid sequence of each subunit with those of other terminal oxidases, especially the Escherichia coli o-type ubiquinol oxidase, which contains heme b, heme o, and one copper ion (5,22,25).To our knowledge, this is the first report on a gene encoding an a1-type cytochrome functioning as a ubiquinol oxidase.MATERIALS AND METHODS Bacterial strains and plasmids. A. aceti 1023 (24) is an isolate from a vinegar fac...

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Isolation and Identification of Acetic Acid Bacteria for Submerged Acetic Acid Fermentation at High Temperature

Cited by 21 publications

References 1 publication

Novel insertion sequence IS1380 from Acetobacter pasteurianus is involved in loss of ethanol-oxidizing ability

Novel insertion sequence IS1380 from Acetobacter pasteurianus is involved in loss of ethanol-oxidizing ability

A new insertion sequence IS1452 from Acetobacter pasteurianus

Characterization of a cytochrome a1 that functions as a ubiquinol oxidase in Acetobacter aceti

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