Proteolytic and lipolytic activities of Micrococcus roseus (65), Halomonas elongata (16) and Vibrio sp. (168) isolated from Danish bacon curing brines

doi:10.1016/0168-1605(94)90136-8

International Journal of Food Microbiology

Volume 22, Issues 2–3, May 1994, Pages 115-126

https://doi.org/10.1016/0168-1605(94)90136-8 Get rights and content

Abstract

Viable cells, cell free extracts and extracellular concentrates of Micorcoccus roseus (65), Halomonas elongata (16) and Vibrio sp. (168) isolated from Danish bacon curing brines were examined for lipase, esterase, proteinase and aminopeptidase activities on natural and synthetic substrates. Micrococcus roseus (65) produced one intracellular esterase with affinity for short chain esters, and two intracellular aminopeptidases with affinity for nonpolar amino acids and l-arginine, respectively. One extracellular esteracellular aminopeptidase with affinity for l-proline was also observed. Three intracellular esterases with affinity for short chain esters and a membrane bound esterase with affinity for butyric to capric esters were found in Halomonas elongata (16), but almost no aminopeptidase acitivity was found. Vibrio sp. (168) had four intracellular esterases with affinity for short chain esters and one esterase with affinity for all tested esters. Furthermore, an enzyme, which did not migrate on electrophoretic gels, had activity on all examined esters and tributyrin. Small intracellular activity on l-alanine was observed for this bacterial strain. There was no proteinase activities in the tested bacteria.

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Comparison of raw hides and correctly cured hides revealed a core microbiome that was absent from contaminated hides. In addition, archaea were missing from well-cured hides, whereas Psychrobacter and Acinetobacter were highly represented (23 % and 17.4 %, respectively). In damaged hides, only a few operational taxonomic units (OTUs), from among the hundreds detected, were able to proliferate and, remarkably, a single Halomonas OTU represented 57.66 % of the reads. Halobacteria, mainly Halovenus, Halorubrum and Halovivax, increased by up to 36.24–39.5 % in the red- and purple-affected hides. The major contaminants were isolated and hide infections, together with collagenase activity, were evaluated. The results showed that hides enriched with the non-pigmented isolate Halomonas utahensis COIN160 damaged the collagen fibers similarly to Halorubrum, and together they were considered to be one of the major causes. Putative degrading inhibitors were also identified from among the Alkalibacillus isolates.
It was concluded that hide contaminations were driven by clonal outbreaks of a few specific microbes, which may have been non-pigmented collagen degraders. Acinetobacter and Alkalibacillus, members of the core microbiome of raw and well-cured salted hides, are suggested as hide contaminant inhibitors that need further analysis.
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Ectoine, a cyclic tetrahydropyrimidine (2-methyl-1,4,5,6-tetrahydropyrimidine-4-carboxylic acid) is a natural compound, which serves as a protective substance in many halophilic actinobacterial cells. Ectoine biosynthesis genes (ectA, B and C) were PCR amplified from the genomic DNA of deep sea actinobacteria, Nocardiopsis alba NIOT-DSB14. The amplified genes were cloned and nucleotide sequences were determined. Sequencing results revealed that ectA, B and C genes contains 471 bp, 1266 bp, 378 bp long ORF encoding 156, 421 and 125 amino acids, respectively. In silico sequence and phylogenetic analysis of nucleotides and amino acids revealed that the ectA, B and C sequences of Nocardiopsis alba NIOT-DSB14 were conserved in many eubacteria.
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Bacterial communities in fish sauce, at different fermentation phases, from two large manufacturing plants (Factories MC and SQ) in Thailand, were studied. The bacterial microbiota in a fish sauce mash of >12 months were examined by 16S rRNA next-generation sequencing. We revealed the presence of 7 phyla, 14 classes, 21 orders, 31 families, and 45 genera of bacteria in the fish sauce. Bacterial communities in the sauces differed across the two factories. In Factory MC, Halanaerobium sp. was the initial dominant genus, which gradually decreased while Lentibacillus sp., Halomonas sp., and Tetragenococcus sp. began to appear in the middle phase. Diversity was the highest in the late fermentation phase. On the contrary, in Factory SQ, high diversity was observed right from the early phase till the late fermentation phase. The bacterial communities varied at different stages of fermentation as well. The most common genera were Peptostreptococcus sp., Peptoniphilus sp., Gallicola sp., Fusobacterium sp., Halanaerobium sp., and Vagococcus sp.. Alpha and beta diversity within the bacterial communities in fish sauce from the factories was also calculated. This is the first report revealing bacterial communities during industrial fermentation of Thai fish sauce. The source of raw materials and bacterial microbiota in the fermentation tank influenced the type of bacteria present throughout the process, and consequently affected the unique product characteristics.
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International Journal of Food Microbiology

Abstract

References (28)

Protease and peptidase activity of Micrococcus species

J. Dairy Sci.

A rapid and sensitive method for the quantification of micrograms quantities of protein utilizing the principles of protein-dye binding

Anal. Biochem.

Kinetics of extracellular protease from the obligately psychrophilic Vibrio OP7 of fish origin

J. Food Prot.

Comparative action of cathepsins D, B, H and L of a new lysosomal cysteine proteinase on rabbit myofibrils

Meat Sci.

Growth profiles of Vibrio species isolated from Danish curing brine

Family II. Vibrionaceae

The unique stability of Vibrio proteolyticus neutral protease under alkaline conditions affords a selective step for purification and use for amino acid-coupling reactions

Appl. Environ. Microbiol.

Purification and characterization of a secreted protease from the pathogenic marine bacterium Vibrio anguillarum

Biochemistry

Extracellular proteinases from Micrococcus GF. II. Isolation and characterization

Lait

One dimensional polyacrylamide gel electrophoresis

Characterization of extracellular alkaline proteases and collagenases induction in Vibrio alginolyticus

J. Gen. Microbiol.

Esterases of lactic Streptococci and their palatability in cheese slime systems

Milchwissenschaft

Judging back bacon packed in bags

Fleischwirtschaft

Microbial catabolism of amino acids during cheese ripening

Sci. Aliments

Cited by (41)

Microbial community on industrial salty bovine hides: From the slaughterhouse to the salting

Combined application of high-throughput sequencing and UHPLC-Q/TOF-MS-based metabolomics in the evaluation of microorganisms and metabolites of dry-cured ham of different origins

Molecular and functional characterization of major compatible solute in Deep Sea halophilic actinobacteria of active volcanic Barren Island, Andaman and Nicobar Islands, India

Next-generation sequencing reveals predominant bacterial communities during fermentation of Thai fish sauce in large manufacturing plants

Ectoine and hydroxyectoine inhibit thermal-induced aggregation and increase thermostability of recombinant human interferon Alfa2b

Food fermentations: Microorganisms with technological beneficial use