WO2020230044A1 - Production d'agents antimicrobiens à partir de déchets végétaux - Google Patents

Production d'agents antimicrobiens à partir de déchets végétaux Download PDF

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WO2020230044A1
WO2020230044A1 PCT/IB2020/054520 IB2020054520W WO2020230044A1 WO 2020230044 A1 WO2020230044 A1 WO 2020230044A1 IB 2020054520 W IB2020054520 W IB 2020054520W WO 2020230044 A1 WO2020230044 A1 WO 2020230044A1
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extract
acid
products
fermentation
concentration comprised
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PCT/IB2020/054520
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English (en)
Inventor
Camilla LAZZI
Valentina BERNINI
Erasmo NEVIANI
Gianni GALAVERNA
Annalisa RICCI
Martina CIRLINI
Antonietta MAOLONI
Luca CALANI
Silvia ZANETTI
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Universita' Degli Studi Di Parma
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Publication of WO2020230044A1 publication Critical patent/WO2020230044A1/fr

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    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
    • C12P7/00—Preparation of oxygen-containing organic compounds
    • C12P7/40—Preparation of oxygen-containing organic compounds containing a carboxyl group including Peroxycarboxylic acids
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • A23L3/00—Preservation of foods or foodstuffs, in general, e.g. pasteurising, sterilising, specially adapted for foods or foodstuffs
    • A23L3/34—Preservation of foods or foodstuffs, in general, e.g. pasteurising, sterilising, specially adapted for foods or foodstuffs by treatment with chemicals
    • A23L3/3454—Preservation of foods or foodstuffs, in general, e.g. pasteurising, sterilising, specially adapted for foods or foodstuffs by treatment with chemicals in the form of liquids or solids
    • A23L3/3463—Organic compounds; Microorganisms; Enzymes
    • A23L3/3472—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 COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • A23L3/00—Preservation of foods or foodstuffs, in general, e.g. pasteurising, sterilising, specially adapted for foods or foodstuffs
    • A23L3/34—Preservation of foods or foodstuffs, in general, e.g. pasteurising, sterilising, specially adapted for foods or foodstuffs by treatment with chemicals
    • A23L3/3454—Preservation of foods or foodstuffs, in general, e.g. pasteurising, sterilising, specially adapted for foods or foodstuffs by treatment with chemicals in the form of liquids or solids
    • A23L3/3463—Organic compounds; Microorganisms; Enzymes
    • A23L3/3481—Organic compounds containing oxygen
    • A23L3/3508—Organic compounds containing oxygen containing carboxyl groups
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • A23L3/00—Preservation of foods or foodstuffs, in general, e.g. pasteurising, sterilising, specially adapted for foods or foodstuffs
    • A23L3/34—Preservation of foods or foodstuffs, in general, e.g. pasteurising, sterilising, specially adapted for foods or foodstuffs by treatment with chemicals
    • A23L3/3454—Preservation of foods or foodstuffs, in general, e.g. pasteurising, sterilising, specially adapted for foods or foodstuffs by treatment with chemicals in the form of liquids or solids
    • A23L3/3463—Organic compounds; Microorganisms; Enzymes
    • A23L3/3526—Organic compounds containing nitrogen
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
    • C12P13/00—Preparation of nitrogen-containing organic compounds
    • C12P13/04—Alpha- or beta- amino acids
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
    • C12P7/00—Preparation of oxygen-containing organic compounds
    • C12P7/02—Preparation of oxygen-containing organic compounds containing a hydroxy group
    • C12P7/22—Preparation of oxygen-containing organic compounds containing a hydroxy group aromatic

Definitions

  • the present invention relates to a process for preparing an extract with antimicrobial activity through the fermentation of by-products or waste products of the fruit and vegetable processing sector.
  • the invention further relates to the extract thus obtained and the use thereof to increase the shelf life of food products.
  • shelf life defined as the interval of time during which a food maintains an acceptable level of safety and quality and thus the period of time that corresponds, under certain storage conditions, to a tolerable decrease in the quality of a food.
  • Shelf life is often also connected with another concept, namely, the marketability of a product.
  • the beginning of processes that modify a food can render it no longer saleable, despite it being still fit for consumption, resulting in a considerable increase in a company’s costs.
  • producers are often faced with the need to make their products more stable in order to cope with distribution chains requiring longer and longer shelf lives, or simply to succeed in preserving the quality of their product better.
  • Foods that are highly perishable from a microbiological viewpoint, for which an expiration date is used are products that must normally be kept in a refrigerator, e.g. fresh cheeses, meat and some meat-based products, fresh pasta, chilled ready meals, fruit and vegetables, etc.
  • the technologies used to improve the preservation and hence the shelf life of food are based essentially on the capacity to limit microbial growth (bacteriostatic effect) or on a definitive inactivation of the associated flora (bactericidal effect).
  • the first group includes, for example, refrigeration or freezing, drying, vacuum preservation or the use of a modified atmosphere and the use of organic or inorganic preservatives. Technologies such as pasteurisation, irradiation with ionising radiation, the application of high hydrostatic pressure (HPP), the use of high-voltage electric charges or the addition of lysozyme, by contrast, bring about a bactericidal effect.
  • HPP high hydrostatic pressure
  • a first aspect of the present invention relates to a process for obtaining an extract from by-products or waste products, preferably of the fruit and vegetable industry.
  • the process of the present invention envisages a step of inoculating the by-products or waste products with at least one bacterium, a fermentation step and a step of extracting the fermented product obtained.
  • lactic bacteria preferably selected from among the species: L. casei, L. paracasei and L. rhamnosus, have proven to be particularly effective.
  • the process of the present invention has shown to be useful for obtaining an extract with antimicrobial capacities.
  • the Applicant has surprisingly found that, from waste products or by products of the fruit and vegetable industry, in particular, by-products of fruit and vegetable processing, one obtains an extract capable of increasing the shelf life of foods by reducing the alterative and pathogenic bioburden.
  • a second aspect of the present invention relates to an extract obtained with the process of the present invention.
  • the extract preferably comprises a mixture of organic acids, polyphenols, peptides and amino acids.
  • a third aspect of the invention relates to the use of the extract obtained with the process of the present invention and described above, or of a composition comprising said extract, as an antimicrobial.
  • a fourth aspect of the present invention relates to the use of the extract obtained with the process of the present invention and described above or of a composition comprising said extract in order to increase the average shelf life of foods.
  • Figure 1 shows a heatmap in which the strains used are represented on the right, along with the incubation times after which the antimicrobial activity was assessed, and at the bottom the pathogenic strains the antimicrobial activity was tested against (different strains of Salmonella, Listeria, E. coli, Staphylococcus aureus, Bacillus cereus). The unfermented sample (control) is indicated at the bottom;
  • Figure 2 shows a graph summarising the main phenolic compounds present in the tomato extract (“PF”) and also present in all three extracts, including the melon and carrot extracts, and the variations in the compounds compared to an unfermented tomato sample (“PC”);
  • Figure 3 shows a graph summarising the main compounds present in the melon extract (“MF”), and also present in the carrot and tomato extracts, and the variations in the compounds compared to an unfermented melon sample (“MC”);
  • Figure 4 shows a graph summarising the main compounds present in the carrot extract (“CF”), and also present in the melon and tomato extracts, and the variations in the compounds compared to an unfermented carrot sample (“CC”);
  • Figure 5 shows the results of a challenge test on raw meat versus various pathogens. It highlights the trend in the presence of the extract of the invention (squares) compared to sodium lactate (diamonds), diacetate (Xs) and meat free of added preservatives (triangles);
  • Figure 6 shows photographs of culture plates inoculated with different pathogens and treated with a solution of organic acids or with the extract fermented with L.rhamnosus 1473.
  • by-product or "waste product” means a secondary product originating from the processing of other products, in particular fruit and vegetable products, normally available at a low cost or free of charge.
  • the terms“by-products” or“waste products” are used interchangeably within the text.
  • the term“active compound” means a substance that possesses a certain biological activity.
  • a first aspect of the present invention relates to a process for preparing a plant extract comprising the steps of:
  • the extract obtained comprises a mixture of active compounds, such as, for example organic acids, polyphenols, peptides and amino acids.
  • the by-products are lyophilised before step (a) for a time comprised from 50 to 80 hours.
  • the by-products are re-hydrated with amounts of water comprised from 50 to 75% w/w, preferably from 60 to 70% w/w.
  • the by-products are then heat treated, preferably in an autoclave, at a temperature higher than 100°C, preferably comprised from 100°C to 130°C, more preferably from 1 10°C and 125°C, for a time comprised from 10 to 40 minutes, preferably comprised from 15 to 30 minutes.
  • the waste products are broken down into parts of a small size, preferably by mechanical destruction, for example by shredding or grinding.
  • the by-products or the waste products are preferably selected from among: cuticles, peels, seeds, pulp, stems and leaves of vegetables or fruit.
  • the by-products or waste products are obtained from fruit selected from among: citrus fruits, preferably oranges, clementines, lemons and mandarins, peaches, apricots, plums, carrots, apples, melons, pears and grapes.
  • the waste by-products are obtained from vegetables selected from among: tomatoes, potatoes, salads, courgettes, carrots, onions, fennel, aubergines, peppers and beans.
  • the by-products or waste products are obtained from the processing of tomatoes, carrots and melon.
  • the bacteria are lactic bacteria, preferably of the genus Lactobacillus, selected from among the species: L. casei, L. paracasei, and L. rhamnosus.
  • the bacteria belong to at least one bacterial strain selected from among: Lactobacillus rhamnosus 1019, Lactobacillus rhamnosus 1473, Lactobacillus paracasei 4186, Lactobacillus casei 2240 and Lactobacillus casei 2246.
  • the strain L. rhamnosus 1019 was deposited in the Belgian Coordinated Collections of Microorganisms (BCCM) on 12 February 2019 with the accession number LMG P-31263 and the following name (designation): UNIPR1019.
  • the strain L. rhamnosus 1019 is characterised by a DNA sequence, preferably a DNA sequence coding for rRNA 16S, said sequence comprising SEQ ID NO: 1 or any sequence characterised by 80-99.9% identity to SEQ ID NO:1 shown in Table 1 .
  • the strain L. rhamnosus 1473 was deposited in the BCCM on 12 February 2019 with the accession number LMG P-31264 and the following name (designation): UNIPR1473.
  • the strain L. rhamnosus 1473 is characterised by a DNA sequence, preferably a DNA sequence coding for rRNA 16S, said sequence comprising SEQ ID NO: 2 or any sequence characterised by 80-99.9% identity to SEQ ID NO:2 L paracasei 4186 was deposited in the BCCM on 12 February 2019 with the accession number LMG P-31266 and the following name (designation): UNIPR4186.
  • the strain L. paracasei 4186 is characterised by a DNA sequence, preferably a DNA sequence coding for rRNA 16S, said sequence comprising SEQ ID NO: 3 or any sequence characterised by 80-99.9% identity to SEQ ID NO:3 L. casei 2240 was deposited in the BCCM on 12 February 2019 with the accession number LMG P-31265 and the following name (designation): UNIPR2240.
  • the strain L. casei 2240 is characterised by a DNA sequence, preferably a DNA sequence coding for rRNA 16S, said sequence comprising SEQ ID NO: 4 or any sequence characterised by 80-99.9% identity to SEQ ID NO:4 L. casei 2246 was deposited in the BCCM on 12 February 2019 with the accession number LMG P-31267 and the following name (designation): UNIPR2246.
  • the strain L casei 2246 is characterised by a DNA sequence, preferably a DNA sequence coding for rRNA 16S, said sequence comprising SEQ ID NO: 5 or any sequence characterised by 80-99.9% identity to SEQ ID NO:5.
  • the bacterial strains used in the process of the present invention are cultured and replicated in vitro before being placed in contact with the by-products or waste products.
  • the bacterial strains are preferably replicated in vitro at least 24 hours before inoculation with the by-products to obtain the fermentation mixture.
  • At least one of the bacterial strains identified above is inoculated with the by-products or waste products to obtain an initial concentration (i.e. prior to fermentation) in the fermentation mixture comprised from 10 4 to 10 11 colony forming units/g (CFU/g), preferably from 10 5 to 10 10 CFU/g, more preferably from 10 7 to 10 8 CFU/g.
  • step (b) of the present process takes place at a temperature comprised from 27°C to 42°C, preferably from 28°C to 39°C, for a time comprised from 48 to 96 hours, preferably comprised from 60 to 80 hours.
  • the concentration of the at least one bacterial strain in the fermented product obtained is comprised from 10 7 to 10 10 CFU/g, preferably from 10 8 and 10 9 CFU/g.
  • the fermented product obtained can be directly subjected to extraction or be preserved by freezing.
  • the fermented product obtained is lyophilised to eliminate most of the water present and to concentrate the sample.
  • the fermented products are preferably lyophilised for 48-96 hours, more preferably for 60-72 hours, before being subjected to extraction according to step c).
  • the fermented product preferably lyophilised, is subjected to at least one extraction in an aqueous and/or organic solvent.
  • the fermented product is subjected to a process of extraction with a water/organic solvent mixture.
  • the organic solvent is preferably an alcohol, for example ethanol or methanol.
  • the aqueous solution further comprises an acid, preferably a carboxylic acid.
  • the extraction with a water/organic solvent mixture also comprises a homogenisation and a centrifugation of the homogenate to recover the liquid phase containing the extract, which can be subsequently filtered.
  • the solid phase recovered by centrifugation can be subjected to a second extraction.
  • the water/organic solvent mixture is water/ethanol in a 30/70 proportion v/v, preferably 40/60 v/v, more preferably 50/50 v/v.
  • Said mixture is preferably acidified with formic acid in a concentration comprised from 0.02 to 0.2%, preferably 0.05 to 0.15% v/v.
  • the extraction solution is then concentrated to give the extract according to the invention, which can be resuspended in a buffer and/or in an aqueous solution.
  • a second aspect of the present invention relates to an extract obtained with the process according to the present invention.
  • the extract comprises a mixture of compounds with antimicrobial activity, preferably with bacteriostatic and/or bactericidal activity.
  • the extract comprises a mixture of compounds selected from among: organic acids, polyphenols, peptides and amino acids.
  • the extract comprises organic acids selected from among: lactic acid, acetic acid, citric acid and combinations thereof.
  • the extract comprises:
  • lactic acid in a concentration comprised from 50 to 90% v/v, preferably from 55 to 85% v/v, more preferably from 60 to 80% v/v;
  • - citric acid in a concentration comprised from 2 to 25% v/v, preferably from 3 to 22% v/v, more preferably from 5 to 20% v/v;
  • - acetic acid in a concentration comprised from 1 to 15% v/v, preferably from 2 to 12% v/v, more preferably from 3 to 10% v/v.
  • the extract comprises organic acids selected from among: lactic acid, acetic acid, succinic acid, glyceric acid, malic acid, tartaric acid, citric acid and a lactic acid dimer.
  • the extract comprises:
  • lactic acid in a concentration comprised from 50 to 90% v/v, preferably from 55 to 85% v/v, more preferably from 60 to 80% v/v;
  • - citric acid in a concentration comprised from 2 to 25% v/v, preferably from 3 to 22% v/v, more preferably from 5 to 20% v/v;
  • - acetic acid in a concentration comprised from 1 to 15% v/v, preferably from 2 to 12% v/v, more preferably from 3 to 10% v/v;
  • - glyceric acid in a concentration comprised from 0.3 to 10% v/v, preferably from 0.5 to 8% v/v, more preferably from 1 to 5% v/v;
  • - malic acid in a concentration comprised from 0.3 to 10% v/v, preferably from 0.5 to 8% v/v, more preferably from 1 to 5% v/v;
  • - lactic acid in dimeric form in a concentration comprised from 0.3 to 10% v/v, preferably from 0.5 to 5% v/v, more preferably from 0.8 to 3% v/v;
  • - succinic acid in a concentration comprised from 0.3 to 10% v/v, preferably from 0.5 to 5% v/v, more preferably from 0.8 to 3% v/v;
  • the extract comprises phenolic compounds selected from among: hydroxyvaleric acid or hydroxyisovaleric acid or methylbutyrrhic acid, leucic acid, phenyllactic acid, caffeic acid, p- hydroxyphenyllactic acid, dihydrocaffeic acid and indole-3-lactic acid.
  • the extract further comprises amino acids and steroidal alkaloids (polyphenols).
  • the amino acids are preferably selected from among: leucine, isoleucine, tryptophan, tyrosine, adenosine and valine.
  • the steroidal alkaloids are preferably selected from among tomatidine, steroidal glycoalkaloid, neorickiioside A/ lycoperoside H and esculeoside B.
  • a third aspect of the invention relates to the use of the extract obtained with the process of the present invention and described above, or of a composition comprising said extract, as an antimicrobial.
  • the extract obtained with the process described in the present invention has in fact shown good antimicrobial activity when assessed with conventional techniques.
  • the extract or the composition comprising the extract is used as an antimicrobial.
  • the extract or the composition is preferably used as an antimicrobial in vitro, for example on products for food use.
  • the extract or the composition which comprises the extract is used as an antibacterial against S. aureus, L. monocytogenes, B. cereus, Salmonella spp and E.coli.
  • the extract obtained from the fermentation of tomato or carrot or melon waste or the composition which comprises it is used as an antibacterial against S. aureus, L monocytogenes, B. cereus, Salmonella spp and E.coli.
  • the extract obtained from the fermentation of melon waste with the strain L. casei 2240 or with L. paracasei 4186 or with L. rhamnosus 1473 or with L. rhamnosus 1019 or the composition which comprises the extract is used as an antibacterial against S. aureus, L. monocytogenes, B. cereus, Salmonella spp and E.coli.
  • the extract obtained from the fermentation of melon waste with the strain L. casei 2240 or the composition which comprises the extract is used as an antibacterial against S. aureus, L. monocytogenes, B. cereus and Salmonella spp.
  • the extract obtained from the fermentation of carrot waste with the strain L. casei 2240 or with L. casei 2246 or with L. paracasei 4186 or with L. rhamnosus 1473 or with L. rhamnosus 1019 or the composition which comprises the extract is used as an antibacterial against S. aureus, L. monocytogenes, B. cereus, Salmonella spp and E.coli.
  • the extract obtained from the fermentation of carrot waste with the strain L. casei 2246 or the composition which comprises the extract is used as an antibacterial against S. aureus, L. monocytogenes, B. cereus, Salmonella spp and E. coli, preferably against B. cereus.
  • the extract obtained from the fermentation of tomato waste or the composition which comprises the extract is used as an antibacterial against L monocytogenes, B. cereus and Salmonella spp.
  • the extract obtained from the fermentation of tomato waste with the strain L. casei 2246 or with L rhamnosus 1473 or with L. rhamnosus 1019 or the composition which comprises the extract is used as an antibacterial against L. monocytogenes, B. cereus and Salmonella spp.
  • the extract or the composition comprising the extract is used as an antifungal, preferably against yeasts.
  • the extract obtained with the process described in the present invention has in fact shown good antifungal activity when assessed with conventional techniques.
  • the extract or the composition is preferably used as an antifungal in vitro, for example, on products for food use.
  • the extract obtained from the fermentation of carrot or melon waste or the composition which comprises it is used as an antifungal, preferably against yeasts.
  • the extract obtained from the fermentation of carrot or melon waste with the strain L. rhamnosus 1019 or the composition which comprises it is used as an antifungal, preferably against yeasts.
  • the tomato, carrot and melon waste was ground, divided into 250 g aliquots, and sterilised in an autoclave at 121 °C for 20 minutes in order to ensure the absence of endogenous microflora in the substrate to be fermented. Afterwards, the sterilised aliquots were stored at - 80°C until the time of use.
  • Microbial strains used for fermentation For fermentation, 7 microbial strains belonging to four different species were used: Lactobacillus casei (2240 and 2246), Lactobacillus paracasei (4186), Lactobacillus rhamnosus (1019 and 1473) and Lactobacillus plantarum (PR1 and PR285).
  • the strains were isolated from plant and dairy matrices (Table 2).
  • strains belong to the collection of the Department of Food Science and Drugs of the University of Parma. All of the strains were kept in stock at -80°C in de Man, Rogosa and Sharpe (MRS) (VWR chemicals, Milan, Italy) supplemented with 12.5% glycerol (v/v).
  • MRS de Man, Rogosa and Sharpe
  • the cultures were revitalized by allowing the strains to grow twice for 24 h, with an inoculation of 3% v/v in MRS broth (VWR) at 37°C (L casei, L. paracasei and L. rhamnosus) and at 30°C ⁇ L. plantarum), under conditions of aerobiosis.
  • VWR MRS broth
  • an inoculation of 3% v/v in MRS broth (VWR) was performed, incubating at 37°C (L. casei, L. paracasei and L. rhamnosus) and at 30 °C (L plantarum) for 15 hours, in order to obtain a cellular concentration of 10 9 CFU/ml.
  • the cells of the overnight culture were collected by centrifugation (10 000 x g, 10 minutes, 4°C), washed twice in Ringer’s solution (VWR chemicals, Milan, Italy), and resuspended in sterile water.
  • the substrates to be fermented were inoculated with an aliquot of the bacterial suspension such as to obtain a final concentration of 10 7 CFU/g.
  • the inoculated substrates were incubated for 72 hours at 37°C (in the case of L casei, L. paracasei and L rhamnosus ) and at 30°C (in the case of L plantarum). Every fermentation was conducted in triplicate.
  • the samples were analysed at the beginning (TO) and end of fermentation (72 hours): 5 g of each sample were supplemented with 45 ml of Ringer’s solution (VWR) and homogenised for 2 minutes at 230 rpm by means of a Stomacher 400 Circulator peristaltic homogeniser (Seward, Worthing, UK).
  • the decimal dilutions of the fermented substrates were prepared in Ringer’s solution (VWR) and plated in MRS agar (VWR); this was followed by incubation at 37°C (L. casei, L. paracasei and L. rhamnosus) and at 30°C (L plantarum) for 72 hours under aerobiosis.
  • the fermented substrates were lyophilised for 72 hours (LIO-5P DIGITAL, Cinquepascal, Ml) in order to eliminate most of the water present and thus increase the concentration of the sample.
  • the lyophilised fermented samples were stored at -80°C until the time of extraction thereof.
  • an extraction process modifying the protocol described by Dall'Asta et al. (2013) was carried out.
  • the solvent used for extraction consisted of water/ethanol 50/50 v/v acidified with 0.1 % formic acid (HCOOH).
  • the solution obtained was filtered through paper to enable recovery of the solid part, to which another 50 ml of solvent were added, followed by a second extraction as previously described.
  • the solution obtained from the second extraction was again filtered through paper for 45 minutes. The two liquid fractions obtained were then united.
  • the solution obtained was filtered through paper to enable recovery of the solid part, to which another 50 ml of solvent were added, followed by a second extraction as previously described.
  • the solution obtained from the second extraction was centrifuged (4000 rpm, 5 minutes, 4°C).
  • the liquid fraction recovered following centrifugation was united with the liquid fraction obtained from the first filtration and the solution obtained was then subjected to vacuum filtration.
  • the extracts were concentrated by using a STRIKE 300 rotary evaporator (Steroglass, Italy, PG). To facilitate the concentration of the extracts, the rotation speed was fixed at 150 rpm and the bath temperature set at 40°C. The concentrated extracts were then resuspended in sterile water so as to obtain a concentration of 60%.
  • the extracts thus obtained were stored at -80°C until the time of use.
  • Antimicrobial activity “agar well diffusion assay”
  • the plates were prepared using Tryptic Soy Agar (TSA) (Oxoid) as the medium for the 14 pathogenic strains and the bulk of potentially alterative microorganisms, and Sabouraud (MAST Group LTD, UK) for the bulk of yeasts.
  • TSA Tryptic Soy Agar
  • MAST Group LTD MAST Group LTD, UK
  • the microbial concentration of the overnight cultures of the pathogenic strains, the bulk of potentially alterative microorganisms and the bulk of yeasts was determined by means of a viable count in TSA (Oxoid), PCA (Oxoid) and YEDC Agar (Remel), respectively.
  • the overnight cultures of the pathogenic strains and the bulk of potentially alterative microorganisms, and the bulk of yeasts were then diluted in Ringer’s solution (VWR) to a concentration of 10 8 CFU/ml.
  • a sterile swab was soaked with each solution obtained as described, at a concentration equal to 10 8 CFU/ml, and used to completely seed the plate containing the agarised medium, on which holes with a diameter of 7 mm were made in order to be filled with 30 pi of each solution to be tested.
  • the antimicrobial activity was assessed for: tomato, melon and carrot extracts, fermented and unfermented, sterile and non-sterile.
  • the plates inoculated with the pathogens were incubated at 37°C, whereas the ones inoculated with the bulk of yeasts and potentially alterative microorganisms were incubated at 30°C.
  • the antimicrobial activity was assessed by measuring the total inhibition halo (mm) observable after 24, 48 and 120 hours of incubation in the case of the pathogenic and alterative microorganisms; after 48 and 120 hours of incubation in the case of yeasts.
  • the unfermented samples i.e. the non-sterile and sterile melon samples
  • the fermented extracts demonstrated greater activity against S. aureus, L monocytogenes and B. cereus, (although the antimicrobial activity versus the latter is considerably reduced over time), while they showed distinctly less activity against E. coli.
  • the extracts that showed the least antimicrobial activity of all were the ones fermented with the two L. plantarum strains, PR1 and PR285, whereas the ones showing the greatest activity belong to the L. casei group, in particular L. casei 2240 and L. paracasei 4186.
  • the extract obtained from fermentation with L. casei 2240 seems to possess the best antimicrobial activity. This activity is very well maintained over time, up to 120 h of observation, against all of the microorganisms tested, with the exception of B. cereus.
  • the extract obtained using L. casei 2240 performed best among all the extracts, especially after 24 h, with an inhibition halo of 6.25 ⁇ 1 .77 mm. However, after 120 h the inhibition halo is reduced to 1 .75 ⁇ 0.88 mm, showing the absolute minimum activity.
  • the extract obtained with L casei 2246 shows its greatest activity against L. monocytogenes after 24 h, with an inhibition halo of 4.56 ⁇ 0.10 mm.
  • this activity is not maintained over time; in particular, it is reduced above all after 120 h (2.03 ⁇ 0.38 mm).
  • This extract also shows good activity against the bulk of alterative microorganisms, which is maintained with the passing of time, whereas it shows the least activity against E. coli, especially after 120 h.
  • the extract deriving from fermentation with L. paracasei 4186 also shows good antimicrobial activity.
  • after 24 h it exhibits its greatest activity versus B. cereus, with an inhibition halo of 6.25 ⁇ 1 .77 mm, which, however, is reduced to 1 .75 ⁇ 0.88 mm at 120 h (absolute minimum activity).
  • the activity against the bulk of alterative microorganisms was maintained over time (mean value 2.89 ⁇ 0.19 mm), revealing a behaviour similar to that of the extract obtained using L. casei 2240.
  • L. rhamnosus 1019 also leads to good activity versus B. cereus at 24 h (4.46 ⁇ 0.18 mm), though it exhibits the same behaviour as the extracts described previously at 120 h.
  • the antimicrobial activity against Salmonella spp by contrast, is very well maintained.
  • the activity against E. coli and S. aureus is maintained as well, at least until 48 h, decreasing after 120 h.
  • S. aureus is the microorganism most inhibited by the extract obtained from fermentation with L. rhamnosus 1473 after 24 h (inhibition halo of 3.86 ⁇ 0.46 mm). Little activity was observed, however, against E. coli, and it decreased further after 120 h, but, contrary to what was observed for E. coli, this extract maintained its activity versus Salmonella spp., B. cereus and the bulk of alterative microorganisms over time. Antimicrobial activity of carrot extracts
  • the unfermented samples i.e. the non-sterile and sterile carrot samples
  • the sterile carrot extracts demonstrate to have greater activity against the two aforementioned microbial species than the non-sterile extracts.
  • the fermented extracts demonstrate greater activity against S. aureus and L monocytogenes, whereas they exhibit distinctly less activity against E. coli.
  • the antimicrobial activity seems to remain constant over time, mainly against the bulk of alterative microorganisms and E. coli.
  • the extracts exhibiting the least antimicrobial activity of all are the ones fermented with L. paracasei 4186, whereas those exhibiting the greatest activity are the ones fermented with strains belonging to the L. casei group, namely, L. casei 2246 and L. rhamnosus 1019.
  • the extract obtained with L. casei 2240 exhibits its maximum activity against S. aureus at 24 h (3.62 ⁇ 0.32 mm) and minimum activity against B. cereus after 120 h (1 .04 ⁇ 0.08 mm).
  • Salmonella spp., E. coli and the bulk of alterative microorganisms show a lower sensitivity to this extract
  • the extract obtained with L. casei 2246 is one of the extracts that exhibits the greatest activity against all of the microorganisms tested, showing maximum activity against B. cereus at 24 h (3.99 ⁇ 0.51 mm) and minimum activity against E. coli after 120 h (1.59 ⁇ 0.57 mm).
  • This extract in addition to having good activity against all of the tested microorganisms, is capable of maintaining it over time, except against B. cereus after 120 h of incubation.
  • the extract obtained with L. paracasei 4186 is the one that has the least antimicrobial activity towards all of the pathogenic strains and the bulk of tested alterative microorganisms; in fact, it exhibits no activity at all against L. monocytogenes, E. coli and B. cereus at 120 h.
  • the 4186 extract shows to be more active against S. aureus, towards which it exhibits a maximum of antimicrobial activity (2.00 ⁇ 0.72 mm) at 24 h, although this activity declines considerably over time.
  • L. rhamnosus 1019 lends the extract good activity against all of the tested pathogenic and alterative microorganisms.
  • the 1019 extract shows the highest activity against S. aureus at 24 h (4.35 ⁇ 1 .03 mm).
  • This extract moreover, demonstrates also to have good activity against L. monocytogenes. Less activity, by contrast, was observed against B. cereus, E. coli, S. aureus and the bulk of alterative microorganisms, although the performances were better as compared to other tested extracts and to the controls.
  • the extract obtained with L. rhamnosus 1473 exhibits fair antimicrobial activity against all of the pathogenic and alterative microorganisms used. The greatest activity was found for S. aureus at 24 h, with an inhibition value of 3.74 ⁇ 0.44 mm; the lowest value of activity was obtained for Salmonella spp. after 120 h, 0.61 ⁇ 0.27 mm. Whereas against both the bulk of alterative microorganisms and E. coli, the activity of the extract is maintained over time.
  • the extracts obtained after fermentation showed greater activity vis-a-vis all of the tested pathogenic microorganisms as compared to those obtained in the absence of fermentation and the two preservatives used, namely, sodium lactate and sodium lactate/sodium diacetate.
  • the most evident results, in terms of the size of the inhibition halo were observed against Salmonella spp., L monocytogenes and B. cereus, whereas less activity was observed against E. coli and S. aureus.
  • the antimicrobial activity of the extracts was observed not only against pathogens, but also against the bulk of alterative microorganisms.
  • the antimicrobial activity manifested does not seem to be correlated with the capacity of replication in this substrate; in fact the two strains 2246 and 1473, which had shown different growth trends, exhibited almost the same inhibition haloes.
  • the antimicrobial activity of the fermented melon and carrot extracts was also tested on a bulk of yeasts isolated from a vegetable product based on melon and thickeners.
  • the antimicrobial activity of the extracts was measured, as for the pathogens, with the “agar well diffusion assay” method, with only two differences: Sabouraud Agar was used as the medium and measurements were performed only at 48 and 120 hours, compatibly with the growth times of yeasts.
  • Both types of extracts, deriving from melon and carrot fermentation show slight activity against the bulk of yeasts isolated from a melon-based vegetable product.
  • the extracts deriving from carrot and melon fermented with L. rhamnosus 1019 are the ones giving the greatest activity against yeasts (Tables 6 and 7); however, it remains much lower than the activity observed against pathogenic microorganisms or alterative bacterial cells (bulk alterative microorganisms).
  • the antimicrobial activity of the fermented extracts was also tested in situ on a melon-based vegetable product.
  • three products were prepared, weighing 1 kg each; 20 g of fermented melon extracts were added to the first, 20 g of fermented carrot extracts to the second and the third product was left as it was so it could be used as a control. Then the growth of alterative microflora present in the vegetable product was monitored by means of a viable plate count at the beginning (TO) and after 7, 15 and 30 days of storage at 4 °C and under vacuum. The analysis was conducted in duplicate. For the preparation with the addition of extracts, use was made of the extracts obtained from fermentation with microorganisms belonging to the L.
  • Concentrations of viable, culturable alterative microorganisms present in the vegetable preparation expressed in Log CFU/g at the initial time (TO ) and after 7, 15 and 30 days. The values are shown as means ⁇ standard deviation.
  • the variation in the pH was also monitored.
  • the pH measurement was performed at the initial time and after 30 days of refrigerated storage under vacuum on samples diluted 1 :10 in Ringer’s solution.
  • the samples supplemented with the fermented extracts at the initial time showed to be more acidic than the control sample (mean value 5.20 ⁇ 0.04) showing a difference from the latter of 1.48.
  • the pH of the samples supplemented with the extracts did not undergo any variations over the 30 days, whereas the pH of the control sample decreased by 1 .17 units (5.51 ⁇ 0.2) (Table 9).
  • the increase in acidity could be due to the metabolism of the endogenous alterative flora, which increases with the passing of days of storage.
  • the antimicrobial activity of the fermented extract was assessed by means of the Microbiological Challenge Test (MCT) in fresh ground pork meat (I), and compared with that in meat with no added extract (II), with the addition of sodium lactate at 2% w/w (III) and of sodium lactate/sodium diacetate (96:4) at 2.5% w/w (IV); each sample was separately inoculated with 5 microbial species: B. cereus, Salmonella spp., E. coli, L. monocytogenes and S. aureus.
  • the concentration of each pathogen was determined at the time of inoculation (TO) and after 3, 6 and 9 days of storage under refrigerated conditions, by means of a viable count in an agarised substrate on Mannitol egg yolk polymyxin agar base-MYP (ISO) (VWR), Rambach Agar (MERCK, Milan, Italy), Chromocult Coliform Agar (MERCK), Listeria selective agar base acc. to Ottaviani and Agosti (ISO) (VWR), and CHROMagar Staph Aureus (CHROMagar, Paris, France), respectively.
  • ISO Mannitol egg yolk polymyxin agar base-MYP
  • MERCK Rambach Agar
  • MERCK Chromocult Coliform Agar
  • MERCK Listeria selective agar base acc. to Ottaviani and Agosti
  • CHROMagar Staph Aureus CHROMagar, Paris, France
  • the strain of L. rhamnosus 1473 increased by 1 .57 Log CFU/g after 72 hours in the substrate used for fermentation, causing a slight acidification of the substrate (from pH 4.26 ⁇ 0.02, before fermentation to 3.82 ⁇ 0.01 after fermentation).
  • the extract demonstrated to possess an antimicrobial activity that was greater than or comparable to that of the traditional preservatives of reference.
  • the antimicrobial action showed to be greater against Salmonella spp. and L monocytogenes, with a decrease in the count of 0.85 and 1 .38 Log CFU/g, respectively, at the end of storage. At the end of storage there was a decrease of 1 .19 Log CFU/g also versus S. aureus, which, however, decreased by 0.88 Log CFU/g in untreated meat.
  • the antimicrobial activity of the extract against B. cereus showed to be comparable to that exhibited by the sodium lactate/sodium diacetate mixture; after 9 days the count of the pathogen was comparable to that of the inoculum, with a growth of 0.40 Log CFU/g. Finally, E. coli showed an initial decrease of 0.36 Log CFU/g, but was subsequently able to grow until reaching a count comparable to that of the inoculum.
  • a complete characterisation of the extract is not yet available, but it may be hypothesised that its antimicrobial activity is due to polyphenols, organic acids and small peptides.
  • the acidity of the extract could have inhibited the growth of the pathogens: various studies have in fact demonstrated the susceptibility of the pathogens used to acidic environments. Jay et al. (2009) reported that the limit pH value allowing the growth of Salmonella can vary depending on the type of acid: 4.4 for lactic, 4.05 for citric and 5.40 for acetic.
  • control C A sample of sterilised unfermented tomato extract (called control C) and a sample of tomato extract fermented with strain 1473 of L rhamnosus (called Ferm_L.r._1473) were analysed by ultra high performance liquid chromatography-electrospray ionization-multiple-stage mass spectrometry (UHPLC-ESI-MSn).
  • Both of the extracts were analysed following dilution (25x) with water acidified with 0.1 % formic acid for analysis with a reversed-phase column, while the extracts were diluted with acetonitrile prior to analysis in the HILIC (hydrophilic interaction liquid chromatography) mode.
  • the samples were analysed by ultra-HPLC interfaced with a linear ion trap mass spectrometer LTQ XL with an H-ESI II source.
  • the carrot and melon extracts were first diluted (factor 10x) with water acidified with 0.1 % formic acid for analysis with a reversed-phase column. In addition to the dilution, both carrot extracts (C and Ferm_L.r._1473) were filtered to remove the particulate that was suspended also after centrifugation. This process, indispensable for the subsequent UHPLC- ESI-MSn analysis, was carried out by means of syringe filters with a nylon membrane ( ⁇ 13 mm, porosity 0.45 pm).
  • the samples were analysed by negative ionisation, with a full-scan data-dependent MS3 method in the range of 100-2000 (m/z) under spectrometric conditions optimised for the analysis of the polyphenol component.
  • the following gradient was used for the analysis in the negative ionisation mode: from 0 to 0.5 minutes 95% B; from 0.5 to 9 minutes solvent B decreased to 49%. Then the percentage of B decreased to 20% so as to enable the chromatography column to be washed, with the organic phase increasing to 80%. After 2 minutes of washing, the gradient was set at the initial conditions (95% B) so as to rebalance the column prior to the subsequent analysis.
  • a further analysis was performed in the positive ionisation mode to analyse glycosteroidal alkaloids by means of a full-scan data-dependent MS3 method in the range of 220-2000 (m/z), using a tomatine standard to optimise the spectrometric conditions.
  • the gradient used was the following: from 0 to 0.5 minutes 80% B; from 0.5 to 8 minutes solvent B decreased to 40%. Then the percentage of B decreased to 20% so as to enable the chromatography column to be washed, with the organic phase increasing to 80%. After 2 minutes of washing, the gradient was set at the initial conditions (80% B) so as to rebalance the column prior to the subsequent analysis.
  • the eluent C was maintained constant at 10% throughout the analysis; only the percentages of A and B were modified.
  • the gradient used was the following: from 0 to 4 minutes it was maintained constant at 89% of A and 1 % of B. From 4 to 14 minutes B increased to 50%; then followed a time for rebalancing the column to the initial conditions so as to rebalance the column for the subsequent analysis.
  • the analysis was performed in the positive ionisation mode by means of a full-scan data-dependent MS3 method in the range of 100- 1000 (m/z)
  • Table 10 Chromatographic and spectrometric information about the phytochemical compounds identified in tomato samples in the negative ionisation mode.
  • the extract was derivatized with silanising agents 5 (hexamethyldisilazane HMDS and trimethylchlorosilane TMCS) and then subsequently analysed by means of the GC-MS technique (gas chromatography coupled to mass spectrometry) with the aim of identifying the compounds present.
  • silanising agents 5 hexamethyldisilazane HMDS and trimethylchlorosilane TMCS
  • GC-MS technique gas chromatography coupled to mass spectrometry
  • Chromatographic separation was obtained in a capillary column with a nonpolar stationary phase (phenyl-methyl-siloxane, 5% phenyl; 30 m x 0.25 mm BP5MS column, 0.25 pm internal film, SGE) by applying a temperature gradient from 60° C to 280°C and using helium as the carrier gas (flow 1 ml/min), whereas the recognition of the substances present 5 was achieved by comparing the mass spectra recorded for each compound (range 41 - 500 m/z) with the mass spectra present in the instrument library (NIST 14). A minimum correspondence of 98% was considered for identification purposes.
  • a nonpolar stationary phase phenyl-methyl-siloxane, 5% phenyl; 30 m x 0.25 mm BP5MS column, 0.25 pm internal film, SGE
  • lactic, acetic, succinic, glyceric, malic, tartaric and citric acid and a lactic acid dimer The acid that is most representative of this fraction of the tomato extract is without a doubt lactic acid (73.50 % of the total), followed by citric acid (1 1 .91 %), acetic acid (5.25 %), glyceric acid (2.64 %), malic acid (2.25 %), lactic acid in dimer 15 form (1 .91 %), succinic acid (1 .84 %) and, finally, tartaric acid (0.69 %).
  • Table 14 Chromatographic and spectrometric information about the phytochemical compounds identified in melon samples in the negative ionisation mode.
  • Table 15 Chromatographic and spectrometric information about the phytochemical compounds identified in carrot samples in the negative ionisation mode.
  • MBC Minimum Bactericidal Concentration
  • MBC minimum concentration
  • the strains were inoculated in TSB supplemented with extract at stepwise concentrations. After 24 hours of incubation at the optimal temperature for the strains (37°C for all with the exception of Pseudomonas, which was incubated at 30°C), a spot was applied on Tryptic Soy Agar (TSA) by drawing 10 mI of every sample.
  • TSA Tryptic Soy Agar
  • the lowest concentration of extract with antimicrobial activity at which no growth of the 17 tested strains was observed on the plate was considered the MBC.
  • MBC data are reported for extracts obtained from the fermentation of by products of:
  • the MBC was influenced by their initial concentration in the medium.
  • Salmonella spp and L. monocytogenes there was an observed tendency of the MBC to decrease in relation to the decrease in the inoculation level.
  • B. cereus was among the most sensitive also in the presence of high contamination.
  • the MBC was not particularly influenced by the initial concentration of the microorganism. Only in the case of 3 strains, when present at the lowest concentration of 2 Log CFU/ml, was a lower MBC obtained.
  • the comparison of the extracts was made vis-a-vis sodium lactate (E325), i.e. the sodium salt of lactic acid, used in pre-packaged preparations of fresh minced meat.
  • concentration of the lactate tested was the one generally used, namely, 2%.
  • the first row indicates the meat sample without the addition of any preservative.
  • the second to fifth rows show the evolution of the microflora in the meat to which the extract was added, in different amounts; the last column indicates the meat to which sodium lactate was added.
  • the extract shows a reduction in alterative microflora, which was not observed in the presence of lactate.
  • FIG. 5 shows some examples of the bactericidal effect of the extracts and other compounds.
  • the squares highlight the evolution of the tested microorganism in the meat in the presence of the extract of the invention, the Xs in the presence of diacetate (preservative used for this product), the diamonds in the presence of lactate (another preservative used); the triangles show the trend of the microorganism in the meat in the absence of added compounds.
  • a characterisation of the extracts was performed by GC-MS and the composition in organic acids was identified.
  • the presence of 8 compounds of interest emerged from the analysis, for example, of tomato extract; in particular, the following were identified: lactic, acetic, succinic, glyceric, malic, tartaric and citric acid and a lactic acid dimer.
  • a solution containing the same amounts of acids as the extract was tested by means of the agar well diffusion assay method. As is shown in Figure 6, the solution of acids does not exhibit antimicrobial activity.

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Abstract

La présente invention concerne un procédé de préparation d'un extrait à activité antimicrobienne à partir de sous-produits ou de déchets de l'industrie des fruits et des légumes. Le procédé comprend une étape de mélange des sous-produits ou des déchets avec au moins un micro-organisme, de préférence une bactérie du genre Lactobacillus, une étape de fermentation et une étape d'extraction ultérieure.
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Cited By (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN112940977A (zh) * 2021-03-05 2021-06-11 中国海洋大学 副干酪乳杆菌Lactobacillus paracasei及其应用
CN114084497A (zh) * 2021-10-25 2022-02-25 中国热带农业科学院海口实验站 一种果蔬保鲜袋及其制备方法与应用
CN115895948A (zh) * 2022-10-24 2023-04-04 浙江一鸣食品股份有限公司 一种高效抑制霉菌生长的副干酪乳杆菌及其培养方法和应用
CN116114710A (zh) * 2023-02-23 2023-05-16 西南大学 一种抑菌剂及其制备方法与应用

Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2012136830A1 (fr) * 2011-04-08 2012-10-11 Chr. Hansen A/S Effet antimicrobien synergique
WO2013153074A1 (fr) * 2012-04-09 2013-10-17 Chr. Hansen A/S Bioprotection faisant appel à des souches de lactobacillus rhamnosus

Patent Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2012136830A1 (fr) * 2011-04-08 2012-10-11 Chr. Hansen A/S Effet antimicrobien synergique
WO2013153074A1 (fr) * 2012-04-09 2013-10-17 Chr. Hansen A/S Bioprotection faisant appel à des souches de lactobacillus rhamnosus

Non-Patent Citations (3)

* Cited by examiner, † Cited by third party
Title
CIRLINI MARTINA ET AL: "Application of lactic acid fermentation to elderberry juice: Changes in acidic and glucidic fractions", LWT- FOOD SCIENCE AND TECHNOLOGY, ACADEMIC PRESS, UNITED KINGDOM, vol. 118, 25 October 2019 (2019-10-25), XP085932982, ISSN: 0023-6438, [retrieved on 20191025], DOI: 10.1016/J.LWT.2019.108779 *
M WADHWA ET AL: "Wastes to worth: value added products from fruit and vegetable wastes.", CAB REVIEWS: PERSPECTIVES IN AGRICULTURE, VETERINARY SCIENCE, NUTRITION AND NATURAL RESOURCES, vol. 10, no. 043, 1 January 2015 (2015-01-01), pages 1 - 25, XP055461537, DOI: 10.1079/PAVSNNR201510043 *
RICCI ANNALISA ET AL: "In vitro metabolism of elderberry juice polyphenols by lactic acid bacteria", FOOD CHEMISTRY, ELSEVIER LTD, NL, vol. 276, 10 October 2018 (2018-10-10), pages 692 - 699, XP085524898, ISSN: 0308-8146, DOI: 10.1016/J.FOODCHEM.2018.10.046 *

Cited By (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN112940977A (zh) * 2021-03-05 2021-06-11 中国海洋大学 副干酪乳杆菌Lactobacillus paracasei及其应用
CN112940977B (zh) * 2021-03-05 2022-03-22 中国海洋大学 副干酪乳杆菌Lactobacillus paracasei及其应用
CN114084497A (zh) * 2021-10-25 2022-02-25 中国热带农业科学院海口实验站 一种果蔬保鲜袋及其制备方法与应用
CN114084497B (zh) * 2021-10-25 2023-05-05 中国热带农业科学院海口实验站 一种果蔬保鲜袋及其制备方法与应用
CN115895948A (zh) * 2022-10-24 2023-04-04 浙江一鸣食品股份有限公司 一种高效抑制霉菌生长的副干酪乳杆菌及其培养方法和应用
CN116114710A (zh) * 2023-02-23 2023-05-16 西南大学 一种抑菌剂及其制备方法与应用

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