Soluble fractions prepared from the strain grown at the exponential and the stationary phases in YPG medium with or without acetic acid were analyzed by two-dimensional gel electrophoresis (Fig. Production of a low‐molecular‐weight, alkaline‐active, thermostable protease by a novel, spiral‐shaped bacterium, Cloning and sequencing of the gene cluster encoding two subunits of membrane‐bound alcohol dehydrogenase from. H. Oxidase: (−) Farr H. Anaerobic growth: (−) As we expected, the transformant carrying multiple copies of the aconitase gene produced 105 g l−1 of acetic acid, whereas the transformant carrying the vector accumulated 84 g l−1 of acetic acid (Fig. exceptions. Uozumi G. Fukaya Horinouchi Acetobacter strains were transformed by the electroporation method [14]. [2], Acetobacter aceti is economically important because it is used in the production of vinegar by converting the ethanol in wine into acetic acid. W. 1982; Takemura et al. A. aceti is considered an acidophile, which means it is able to survive in acidic environments, due to having an acidified cytoplasm which makes nearly all proteins in the genome to evolve acid stability. (. Ampicillin was used at a final concentration of 100 μg ml−1 when necessary to maintain plasmids. The above characteristics make them involved in the production of fermented foods, either in a beneficial (chocolate products, coffee, vinegar, and specialty beers) or in a … T. Escherichia coli was routinely cultured in LB medium (10 g l−1 bacto tryptone, 5 g l−1 yeast extract, and 10 g l−1 NaCl; pH 7.0). Ohmori et al. 5. Acetic acid bacteria (AAB) are capable of oxidizing ethanol as substrate to produce acetic acid in neutral and acidic media under aerobic conditions. DNA–DNA hybridization for cloning the gene was performed by the standard method [15, 16] with a Hybond-N+ nylon membrane (Amersham Biosciences). Kaspar Nucleotide sequence was determined by the dideoxy chain termination method [17] combined with the M13 cloning system [18] on a Shimadzu DSQ1000 DNA sequencer. (, Fukaya The NH2-terminal amino acid sequence determined with the protein blotted on a PVDF membrane is present in the deduced amino acid sequence at positions 1–15. Recombinant Microbes for Industrial and Agricultural Applications. Resistance to acetic acid does not always result from resistance to low pH, since bacterial strains capable of growing at low pHs cannot grow when adjusted with acetic acid. AAB are typically considered to … Because the consensus promoter sequence of Acetobacter has not been elucidated, we placed the aconitase gene downstream of the E. coli lac promoter in the E. coli–Acetobacter shuttle vector plasmid, pMV24, whose copy number in A. aceti was estimated to be about 10 [24]. They are Gram-negative, acidophilic α-proteobacteria, and widespread in nature. E. coli was transformed as described by Hanahan [13]. Horinouchi S. R.D. N.J. It furthers the University's objective of excellence in research, scholarship, and education by publishing worldwide, This PDF is available to Subscribers Only. Acetic acid bacteria (AAB) are a group of Gram-negative bacteria which oxidize sugars or ethanol and produce acetic acid during fermentation. The third position of each triplet was designed to be inosine. Amplification of the aconitase gene by use of a multicopy plasmid in A. aceti enhanced the enzymatic activity and acetic acid resistance. For a long time it has been used in the fermentation industry to produce acetic acid from alcohol. III. Isolation and identification of acetic acid bacteria for submerged acetic acid fermentation at high temperature. Influence of the final ethanol concentration on the acetification and production rate in the wine vinegar process. H. The nucleotide sequences of the aconitase gene has been deposited to the DDBJ, EMBL and GenBank databases under Accession No. T. Y. Takemura The soluble fractions from A. aceti 10–8S2, which was grown in the medium without (a and c) or with 1% (v v−1) acetic acid (b and d), were prepared as described in Section 2. The reasons for thermostability of thermophilic strains are still uncertain. The effects of temperature on gluconic acid production and cell growth were coincident in trend with acetic acid formation. Please check your email for instructions on resetting your password. (, Altschul It lives wherever sugar fermentation occurs. (a) The A. aceti transformant was cultured in the presence of 0% (circle), 1% (v v−1) (triangle), 1.75% (diamond), and 2.25% (square) acetic acid. The plasmid was stably maintained during the cultivation periods. A.L. The strains were cultured for 6 h at 37 °C in LB medium (pH 7.0) with or without 1 mM IPTG and 0.05% (vv−1) acetic acid (pH 5.0). T. J. Acetobacter aceti is a Gram-negative bacterium that moves using its peritrichous flagella. The A. aceti transformant was cultured at 400 rpm under 0.2 vvm aeration at 30 °C in YPG medium with constant feeding of ethanol at 1% (v v−1) in a jar-fermentor, as described by Section 2. In this study, Elevation of aconitase activity through gene amplification was expected to enhance acetic acid resistance. Acetate oxidation: (, All strains were tested in a medium containing 5% (v/v) ethanol and incubated at 30 °C with shaking at 150 rev min. The cellular fatty acid composition in acetic acid bacteria. M. Amikam However, some evidence indicates it can be harmful to plants and other flora, though it exists naturally in the world. Okumura A. International Journal of Systematic and Evolutionary Microbiology. 4(b)). [1], "Acetobacter aceti Final Risk Assessment – Biotechnology Program Under Toxic Substances Control Act (TSCA) – US EPA",, Creative Commons Attribution-ShareAlike License, This page was last edited on 15 June 2019, at 12:17. Although the transformant carrying the vector was unable to grow in the medium containing 2.25% (v v−1) acetic acid, the transformant carrying pACO300 could grow in the same medium (Fig. C. 5). Acetic acid was produced by Acetobacter sp. Aconitase activity of the transformant harboring pACO300 at the acetic acid concentration of 60 g l−1 was 0.054 U mg−1 of protein, which was higher than that of the transformant harboring the vector (0.036 U mg−1 of protein), although at the stationary phase at the acetic acid concentration of higher than 80 g l−1 the activity was not detected in both cultures. T. Growth in the presence of 0.35% acetic acid has been used as a criterion; however, while members of the genera Acetobacter, Gluconobacter, Gluconacetobacter,andKomagataeibacter grow in the presence of 0.35% acetic acid, not all genera can, for example, Asaia. Schwerdel


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