Classifications

• • A—HUMAN NECESSITIES
  • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
  • A23B—PRESERVATION OF FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES; CHEMICAL RIPENING OF FRUIT OR VEGETABLES
  • A23B7/00—Preservation of fruit or vegetables; Chemical ripening of fruit or vegetables
  • A23B7/10—Preserving with acids; Acid fermentation
• • A—HUMAN NECESSITIES
  • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
  • A23L—FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES, NOT OTHERWISE PROVIDED FOR; PREPARATION OR TREATMENT THEREOF
  • A23L19/00—Products from fruits or vegetables; Preparation or treatment thereof
  • A23L19/20—Products from fruits or vegetables; Preparation or treatment thereof by pickling, e.g. sauerkraut or pickles
• • A—HUMAN NECESSITIES
  • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
  • A23B—PRESERVATION OF FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES; CHEMICAL RIPENING OF FRUIT OR VEGETABLES
  • A23B2/00—Preservation of foods or foodstuffs, in general
  • A23B2/70—Preservation of foods or foodstuffs, in general by treatment with chemicals
  • A23B2/725—Preservation of foods or foodstuffs, in general by treatment with chemicals in the form of liquids or solids
  • A23B2/729—Organic compounds; Microorganisms; Enzymes
  • A23B2/733—Compounds of undetermined constitution obtained from animals or plants
• • A—HUMAN NECESSITIES
  • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
  • A23L—FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES, NOT OTHERWISE PROVIDED FOR; PREPARATION OR TREATMENT THEREOF
  • A23L29/00—Foods or foodstuffs containing additives; Preparation or treatment thereof
  • A23L29/065—Microorganisms
• • A—HUMAN NECESSITIES
  • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
  • A23L—FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES, NOT OTHERWISE PROVIDED FOR; PREPARATION OR TREATMENT THEREOF
  • A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
  • A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
  • A23L33/105—Plant extracts, their artificial duplicates or their derivatives
• • C—CHEMISTRY; METALLURGY
  • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
  • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
  • C12N1/00—Microorganisms; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
  • C12N1/20—Bacteria; Culture media therefor
• • C—CHEMISTRY; METALLURGY
  • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
  • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
  • C12N1/00—Microorganisms; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
  • C12N1/20—Bacteria; Culture media therefor
  • C12N1/205—Bacterial isolates
• • A—HUMAN NECESSITIES
  • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
  • A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
  • A23V2400/00—Lactic or propionic acid bacteria
  • A23V2400/11—Lactobacillus
  • A23V2400/179—Sakei
• • C—CHEMISTRY; METALLURGY
  • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
  • C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
  • C12R2001/00—Microorganisms ; Processes using microorganisms
  • C12R2001/01—Bacteria or Actinomycetales ; using bacteria or Actinomycetales
  • C12R2001/225—Lactobacillus

Definitions

• This invention relates to fermenting of plant-derived material containing glucosinolates with a starter containing a lactic acid bacterium, as well as to the lactic acid bacterium in question.
• Degradation products of the glucosinolates produced by the bacterium exhibit among other things antimicrobial properties, and may possess a health-promoting effect.
• lactic acid bacteria Preservation of vegetables by fermenting, utilizing lactic acid bacteria is an old process.
• the health-promoting effect of fermented products may at least partially be associated with intestinal function.
• the balancing effect of probiotic lactic acid bacteria on disturbed intestinal microbial flora, among other things, is well known (Gorbach, 1990), and probi- otics are believed to lower serum cholesterol, improve the immune response and alleviate allergies.
• lactic acid bacteria decrease the activity of enzymes that activate carcinogenic compounds (Sanders, 1998, Ziemer and Gibson, 1998).
• glucosinolates and their degradation products occurring in cabbage plants have been shown to exhibit an anticarcinogenic effect (Nastruzzi et al., 1996, Leoni et al., 1997, Zhang and Talalay, 1994).
• Degradation products of glucosinolates have been shown to inhibit the growth of Escherichia coli (Brabban and Edvards, 1995) and the yeasts Saccharomyces cerevisiae, Torulopsis etchellsii, Hansenula rnrakii, Pichia membranefaciens (Kyung and Fleming, 1997).
• Glucosinolates are sulphur- and nitrogen- containing secondary plant compounds consisting of amino acid and sugar. The significance of glucosinolates is still unknown, but they have, among other things, been found to participate in the regulation of sulphur balance, germination and plant defence (Keski- talo, 2001). Glucosinolate-containing cruciferous plants can be used to clear up soil of herbicides. Glucosinolates are intrinsically non-toxic, but compounds formed from them may in high concentrations be detrimental to living organisms (Jaakkola, 2001).
• Benefits of these compounds as compared with pesticides are their spreading to the soil from plant roots and with plant litter and their rapid decomposition in the soil. They therefore are well suited for biological plant protection (Jaakkola, 2001). Glucosinolates and their degradation products occurring in cabbage plants are important
• glucosinolates are decomposed by the enzyme myrosinase, and methods of processing affect the amounts of degradation products (de Vos and Blijleven, 1988, Slominski and Campbell, 1989, S ⁇ rensen, 1990, Rosa and Heaney, 1993, Yen and Wei, 1993, Verkerk et al, 1997).
• glucosinolates are decomposed within two weeks (Daxenbichler et al, 1980, Tolonen et al, 2002).
• the present invention relates to a starter that is applicable to fermenting of plant material, comprising the lactic acid bacterium Lactobacillus sp. 96/142 isolated from sauerkraut.
• the starter of the invention is mainly aimed at fermenting of cabbage plants.
• the invention further relates to the above-mentioned Lactobacillus strain, which is likely to belong to the species Lactobacillus sakei.
• the Lactobacillus sp. 96/142 strain of the invention produces from available sugars via fermentation among other things lactic acid, whereby growth of spoilage microorganisms is prevented. It additionally produces degradation products of glucosinolates possessing inhibitory effects on microbial growth and potential anticarcinogenic activity.
• Probiotic properties of the strain Lactobacillus sp. 96/142 were investigated in in vitro experiments and the strain was found to remain well viable at pH 4 but not at pH 2. The strain is thus expected to remain viable in the intestine, especially if it has protected itself with a self- produced exopolysaccharide or if it is microcapsulated.
• the health-promoting effect of the strain in the intestine is among other things based on its ability to produce from glu- cosinolates degradation products that may possess anticarcinogenic effects.
• production of lactic acid and lowering of intestinal pH are well-known properties balancing the intestinal flora.
• the starter is added to the material to be fermented either as a freeze-dried powder or pre- cultured in vegetable juice.
• other lactic acid bacterial strains can be added that affect the sensory and hygienic quality of the product but do not prevent the glucosinolate-degrading activity of the lactic acid bacterial strain of the invention.
• the culturing temperature should remain within the range 18 - 35 °C, as the strain of the invention is mesophilic.
• the material to be fermented can be fresh or heat-treated plant material. By the heat treatment the plant's own myrosinase activity can almost completely be inactivated without a total destruction of glucosinolates (Ryhanen et al, 2001).
• the material is usually supplemented with salt, which by means of osmotic pressure removes fluid and nutrients from the plant cell tissue for microbial growth.
• Figures 1.1 and 1.2 present the experimental series 1 carried out with head cabbage during years 2000 - 2001 : 1.1.
• Figures 2.1 and 2.2 present fermentation experiments with cauliflower
• Figures 3(a) to 3(f) present the growth of indicator strains E. coli, L. monocytogenes and C. lambica in fermented cabbage juice.
• the change of the optical density of the cell mass is shown in absorbance (A ⁇ Qo) against time on a microtitre plate.
• E. coli (3 a, 3b); L. monocytogenes (3c, 3d) and C. lambica (3e, 3f).
• Lactobacillus sp. 96/142 was deposited on 22 November 2002 according to the Budapest treaty in the DSMZ depository (Deutsche Sammlung von Mikroorganismen und Zellkulturen, • GmbH) under the accession number DSM 15298.
• Lactobacillus sp.2* 12.5 ⁇ 0.2 1.0 ⁇ 0.1 11.5 ⁇ 0.3 0.5 ⁇ 0.1 2.5 x 10 9
• Cabbages fermented with the starter were of equal quality, whereas the quality varied among spontaneously fermented cabbages.
• the number of yeasts and moulds ranged from 100 to 1000 cfu/ml and that of coliform enterobacteria from 0 to 100 cfu/ml. Their growth was prevented when the sauerkaut was fermented with pure cultures of lactic acid bacteria.
• the numbers of yeasts and moulds ranged from 0 to 10 cfu/ml and those of coliform enterobacteria from 0 to 1000 cfu/ml.
• the highest sensory quality was obtained with Leuconostoc sp.78 (DSM 14486) and with Lactobacillus sp.2* (DSM 14485).
• glucosinolate concentrations of head cabbage were determined dur- ing four years (Table 3). Table 3. Concentrations of glucosinolates in cabbage during years 1998 - 2000 (Tolonen et ah, 2002) and 2001 (unpublished) ( ⁇ mol/g d.m.).
• Volatile degradation products of glucosinolates in sauerkrauts fermented with pure cultures are shown in Figures 1.1 and 1.2.
• the total concentration of degradation products was 2 to 3 times higher in the sauerkraut fermented with strain Lactobacillus sp. 96/142 as compared with the others.
• concentrations of 3-methylsulfinyl ⁇ ropyl-ITC and allyl-ITC were multifold with strain Lactobacillus sp. 96/142.
• Concentrations of indole carbinol and indole acetonitrile were below detection limit (10 - 50 ⁇ g/lOOg f.wt.) in all fermentations.
• Indole glucosinolates may occur as ascorbigen, but they were not determinend.
• the amount of degradation products of glucosinolates is related with the amount of starting raw material. Concentration of goitrin was high in all treatments (500 - 1100 ⁇ g/100 g f.wt.).
• Head cabbages (variety Lennox) had been grown at MTT Horticulture with nitrogen sup- plementation (A) and without nitrogen supplementation (B) during the growing season.
• the cabbages were fermented by adding the strains Leuconostoc sp. 78, Lactobacillus sp. 2* and Lactobacillus sp. 96/142 at 10 5 - 10 6 cfu/g.
• the cabbages were fermented for 14 d. pH and amount of glucose were higher in the sauerkraut prepared from cabbages grown without nitrogen supplementation than those of cabbages fertilized during the growing season, and the pH and sugar of sauerkrauts prepared with strain Lactobacillus sp.
• the cauliflower experiment A preservation experiment with comme ⁇ cial cauliflower was carried out on 24.8.2001 with Lactobacillus sp. 96/142 and Leuconostoc sp. 78. Grated cauliflower was supplemented with 1 % of table salt and starter at 10 s cfu/g and fermenting was continued for 3 d. In both treatments the growth of coliform enterobacteria was prevented in three days and the pH decreased to 3.9 - 4.0. Lactobacillus sp. 96/142 produced an aromatic and atypical taste to the cauliflower, deviating form the savoury and fresh cauliflower prepared with Leuconostoc sp. 78.
• microorganisms The following microorganism was deposited according to the Budapest treaty in the DSMZ depository (Deutsche Sammlung von Mikroorganismen und Zellkulturen, GmbH, Mascheroder Weg lb, D-38124 Braunschweig, Germany)
• Keskitalo M. 2001. Glukosinolaattien biokemia ja esiintymiseen vaikuttavat kasvuteki- jat.
• Kasviperaiset biomolekyylit - glukosinolaatit ed. Hyvarinen, H.

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• Chemical Kinetics & Catalysis (AREA)
• Botany (AREA)
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• Micro-Organisms Or Cultivation Processes Thereof (AREA)
• Coloring Foods And Improving Nutritive Qualities (AREA)
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• 2003
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• 2004
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