Screening of Antagonistic/Synergistic Effect between Lactic Acid Bacteria (LAB) and Yeast Strains Isolated from Kefir

Kefir is a traditional fermented refreshing beverage which is known for its valuable and beneficial properties for human health. Mainly yeast species, lactic acid bacteria (LAB) strains and fewer acetic acid bacteria strains live together in a natural matrix named “kefir grain”, which is formed from various proteins and polysaccharides. Different microbial species live together in slimy kefir grain and it has been thought that synergetic effect could take place between microorganisms, which belong to different genera and species. In this research, yeast and LAB were isolated from kefir samples obtained from Uludag University Food Engineering Department. The cell morphology of isolates was screened by microscopic examination. Gram reactions of bacteria isolates were determined by Gram staining method, and as well catalase activity was examined. After observing the microscopic/morphological and physical, enzymatic properties of all isolates, they were divided into the groups as LAB and/or yeast according to their physicochemical responses to the applied examinations. As part of this research, the antagonistic/synergistic efficacy of the identified five LAB and five yeast strains to each other were determined individually by disk diffusion method. The antagonistic or synergistic effect is one of the most important properties in a co-culture system that different microorganisms are living together. The synergistic effect should be promoted, whereas the antagonistic effect is prevented to provide effective culture for fermentation of kefir. The aim of this study was to determine microbial interactions between identified yeast and LAB strains, and whether their effect is antagonistic or synergistic. Thus, if there is a strain which inhibits or retards the growth of other strains found in Kefir microflora, this circumstance shows the presence of antagonistic effect in the medium. Such negative influence should be prevented, whereas the microorganisms which have synergistic effect on each other should be promoted by combining them in kefir grain. Standardisation is the most desired property for industrial production. Each microorganism found in the microbial flora of a kefir grain should be identified individually. The members of the microbial community found in the glue-like kefir grain may be redesigned as a starter culture regarding efficacy of each microorganism to another in kefir processing. The main aim of this research was to shed light on more effective production of kefir grain and to contribute a standardisation of kefir processing in the food industry.

Production of Apricot Vinegar Using an Isolated Acetobacter Strain from Iranian Apricot

Vinegar or sour wine is a product of alcoholic and subsequent acetous fermentation of sugary precursors derived from several fruits or starchy substrates. This delicious food additive and supplement contains not less than 4 grams of acetic acid in 100 cubic centimeters at 20°C. Among the large number of bacteria that are able to produce acetic acid, only few genera are used in vinegar industry most significant of which are Acetobacter and Gluconobacter. In this research we isolated and identified an Acetobacter strain from Iranian apricot, a very delicious and sensitive summer fruit to decay, we gathered from fruit's stores in Isfahan, Iran. The main culture media we used were Carr, GYC, Frateur and an industrial medium for vinegar production. We isolated this strain using a novel miniature fermentor we made at Pars Yeema Biotechnologists Co., Isfahan Science and Technology Town (ISTT), Isfahan, Iran. The microscopic examinations of isolated strain from Iranian apricot showed gram negative rods to cocobacilli. Their catalase reaction was positive and oxidase reaction was negative and could ferment ethanol to acetic acid. Also it showed an acceptable growth in 5%, 7% and 9% ethanol concentrations at 30°C using modified Carr media after 24, 48 and 96 hours incubation respectively. According to its tolerance against high concentrations of ethanol after four days incubation and its high acetic acid production, 8.53%, after 144 hours, this strain could be considered as a suitable industrial strain for a production of a new type of vinegar, apricot vinegar, with a new and delicious taste. In conclusion this is the first report of isolation and identification of an Acetobacter strain from Iranian apricot with a very good tolerance against high ethanol concentrations as well as high acetic acid productivity in an acceptable incubation period of time industrially. This strain could be used in vinegar industry to convert apricot spoilage to a beneficiary product and mentioned characteristics have made it as an amenable strain in food and agricultural biotechnology.

Characterization of an Acetobacter Strain Isolated from Iranian Peach that Tolerates High Temperatures and Ethanol Concentrations

Vinegar is a precious food additive and complement as well as effective preservative against food spoilage. Recently traditional vinegar production has been improved using various natural substrates and fruits such as grape, palm, cherry, coconut, date, sugarcane, rice and balsam. These neoclassical fermentations resulted in several vinegar types with different tastes, fragrances and nutritional values because of applying various acetic acid bacteria as starters. Acetic acid bacteria include genera Acetobacter, Gluconacetobacter and Gluconobacter according to latest edition of Bergy-s Manual of Systematic Bacteriology that classifies genera on the basis of their 16s RNA differences. Acetobacter spp as the main vinegar starters belong to family Acetobacteraceae that are gram negative obligate aerobes, chemoorganotrophic bacilli that are oxidase negative and oxidize ethanol to acetic acid. In this research we isolated and identified a native Acetobacter strain with high acetic acid productivity and tolerance against high ethanol concentrations from Iranian peach as a summer delicious fruit that is very susceptible to food spoilage and decay. We used selective and specific laboratorial culture media such as Standard GYC, Frateur and Carr medium. Also we used a new industrial culture medium and a miniature fermentor with a new aeration system innovated by Pars Yeema Biotechnologists Co., Isfahan Science and Technology Town (ISTT), Isfahan, Iran. The isolated strain was successfully cultivated in modified Carr media with 2.5% and 5% ethanol simultaneously in high temperatures, 34 - 40º C after 96 hours of incubation period. We showed that the increase of ethanol concentration resulted in rising of strain sensitivity to high temperature. In conclusion we isolated and characterized a new Acetobacter strain from Iranian peach that could be considered as a potential strain for production of a new vinegar type, peach vinegar, with a delicious taste and advantageous nutritional value in food biotechnology and industrial microbiology.

Isolation and Identification of an Acetobacter Strain from Iranian White-Red Cherry with High Acetic Acid Productivity as a Potential Strain for Cherry Vinegar Production in Foodand Agriculture Biotechnology

According to FDA (Food and Drug Administration of the United States), vinegar is definedas a sour liquid containing at least 4 grams acetic acid in 100 cubic centimeter (4% solution of acetic acid) of solution that is produced from sugary materials by alcoholic fermentation. In the base of microbial starters, vinegars could be contained of more than 50 types of volatile and aromatic substances that responsible for their sweet taste and smelling. Recently the vinegar industry has a great proportion in agriculture, food and microbial biotechnology. The acetic acid bacteria are from the family Acetobacteraceae. Regarding to the latest version of Bergy-s Mannual of Systematic Bacteriology that has categorized bacteria in the base of their 16s RNA differences, the most important acetic acid genera are included Acetobacter (genus I), Gluconacetobacter (genus VIII) and Gluconobacter (genus IX). The genus Acetobacter that is primarily used in vinegar manufacturing plants is a gram negative, obligate aerobe coccus or rod shaped bacterium with the size 0.6 - 0.8 X 1.0 - 4.0 μm, nonmotile or motile with peritrichous flagella and catalase positive – oxidase negative biochemically. Some strains are overoxidizer that could convert acetic acid to carbon dioxide and water.In this research one Acetobacter native strain with high acetic acid productivity was isolated from Iranian white – red cherry. We used two specific culture media include Carr medium [yeast extract, 3%; ethanol, 2% (v/v); bromocresol green, 0.002%; agar, 2% and distilled water, 1000 ml], Frateur medium [yeast extract, 10 g/l; CaCO3, 20 g/l; ethanol, 20 g/l; agar, 20 g/l and distilled water, 1000 ml] and an industrial culture medium. In addition to high acetic acid production and high growth rate, this strain had a good tolerance against ethanol concentration that was examined using modified Carr media with 5%, 7% and 9% ethanol concentrations. While the industrial strains of acetic acid bacteria grow in the thermal range of 28 – 30 °C, this strain was adapted for growth in 34 – 36 °C after 96 hours incubation period. These dramatic characteristics suggest a potential biotechnological strain in production of cherry vinegar with a sweet smell and different nutritional properties in comparison to recent vinegar types. The lack of growth after 24, 48 and 72 hours incubation at 34 – 36 °C and the growth after 96 hours indicates a good and fast thermal flexibility of this strain as a significant characteristic of biotechnological and industrial strains.