Abstract
The importance of the study of acetic bacteria, on species of the Gluconobacter genus is based on its industrial application, as these possess the capacity of bioconversion of sorbitol to sorbose, enabling the process of vitamin C production. The study involved samples collected in industries of soft drinks, flowers, fruits and honey, followed by purification, phenotypic identification, molecular identification with the use of primer defined from Nucleotide Sequence Database consultation. Strains preserved were identified as members of the Acetobacteraceae family, Gluconobacter genus. 110 strains had been isolated of substrate: Pyrostegia venusta (ker-gawler), honey, Vitis vinifera (grape), Pyrus communis (pear), Malus sp. (apple) and in two samples of soft drinks. Of this total 57 strains had been recovered in manitol medium (manitol, yeast extract, peptone), 12 in YMG medium (glucose, manitol, yeast extract, ethanol, acetic acid), 41 in enrichment medium (De Ley and Swings) and later in the GYC medium (glucose, yeast extract and calcium carbonate). 68 strains were identified as Gram negative bacilli rods. Of these, 31 were characterized biochemically as belonging to the Acetobacteriaceae family as they were catalase positive, oxidase negative and producers of acid from glucose. The characterization of these strains was complemented with the biochemistry tests: gelatin liquefaction, nitrate reduction, indole and H2S production, oxidation of ethanol to acetic acid and molecular tests for genus identification. Only eight strains were characterized as pertaining to the Gluconobacter genus. The strains are maintained in collection cultures at the Microbiology Laboratory of the Biology Department at the Sao Paulo State University (UNESP) in Assis, stored in malt extract at -196 oC.
Highlights
The importance of the study of acetic bacteria, on species of the Gluconobacter genus is based on its industrial application, as these possess the capacity of bioconversion of sorbitol to sorbose, enabling the process of vitamin C production
Santos (2000) realizou um trabalho de isolamento, purificação, identificação rápida e armazenamento de bactérias acéticas isoladas em fermentadores de vinagre, entre as quais algumas cepas foram identificadas como pertencentes ao gênero Gluconobacter
A extração do DNA genômico das linhagens selecionadas foi realizada por meio do uso de tampão de lise contendo CTAB (2% p/v de brometo de acetiltrimetilamônio – CTAB – e 1.4 M NaCl; 0,2% v/v de β-mercapto etanol; 20 mM EDTA; 100 mM Tris HCl pH 8), e 3 etapas utilizando: fenol, fenol:clorofórmio (1:1) e clorofórmio: álcool isoamílico (24:1)
Summary
O gênero Gluconobacter pertence à família Acetobacteriaceae. É constituído por bactérias que possuem forma de bastonetes, Gram-negativos e não formam endósporos. As bactérias acéticas constituem um grupo com importantes aplicações industriais, particularmente as do gênero Gluconobacter, que podem ser usadas para conversão oxidativa de sorbitol a sorbose, um intermediário importante na produção (química) do ácido L-ascórbico (vitamina C). Santos (2000) realizou um trabalho de isolamento, purificação, identificação rápida e armazenamento de bactérias acéticas isoladas em fermentadores de vinagre, entre as quais algumas cepas foram identificadas como pertencentes ao gênero Gluconobacter. O presente trabalho teve como objetivo instituir um banco de microrganismos que apresentassem as características: bastonetes, Gram negativos, oxidase negativos, catalase positivos, capazes de acidificar glicose, que não liquefizessem gelatina, não formassem indol a partir do triptofano e que pudessem ser identificados como pertencentes à família Acetobacteriaceae, gênero Gluconobacter complementados com análise de caracterização molecular pela metodologia de RAPD-PCR
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