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G Corrieu

Publications and source records attributed to G Corrieu.

At least 37 records · Page 2Linked to original sources

Diacetyl and alpha-acetolactate overproduction by Lactococcus lactis subsp. lactis biovar diacetylactis mutants that are deficient in alpha-acetolactate decarboxylase and have a low lactate dehydrogenase activity.

Lactococcus lactis subsp. lactis biovar diacetylactis strains are utilized in several industrial processes for producing the flavoring compound diacetyl or its precursor alpha-acetolactate. Using random mutagenesis with nitrosoguanidine, we selected mutants that were deficient in alpha-acetolactate decarboxylase and had low lactate dehydrogenase activity. The mutants produced large amounts of alpha-acetolactate in anaerobic milk cultures but not in aerobic cultures, except when the medium was supplemented with catalase, yeast extract, or hemoglobin.

Aerobiosis↗

Behavior of Brevibacterium linens and Debaryomyces hansenii as ripening flora in controlled production of smear soft cheese from reconstituted milk: growth and substrate consumption dairy foods.

Experimental cheeses inoculated with Debaryomyces hansenii and Brevibacterium linens were ripened for 76 d under aseptic conditions. Triplicate cheese-making trials were similar as a result of efficient control of the atmosphere. In all trials, D. hansenii grew rapidly during the first 2 d and then slowed, but growth remained exponential until d 10 (generation time around 70 h). Total cell counts were higher than the number of viable cells, and after 10 d they remained around 3 x 10(9) yeast/g of DM. This difference resulted from the nonviability of a fraction of D. hansenii. After d 15, the pH of the rind was close to 7, and B. linens grew exponentially until d 25 (generation time around 70 h). The growth rate subsequently decreased but remained exponential (generation time around 21 d). Cell counts of D. hansenii and B. linens were correlated with the environmental technical conditions. Total D. hansenii counts were also correlated with total B. linens counts. Viable B. linens counts were related to rind lactate, and total counts depended on rind pH, internal lactate, and D. hansenii viable counts. The internal pH of the cheese depended on lactate concentrations, whereas surface pH was related to internal lactose, temperature, and relative humidity. These results suggest a determining role of the diffusion of the carbon sources in the ripening of smear soft cheese.

Atmosphere Exposure Chambers↗

Behavior of Brevibacterium linens and Debaryomyces hansenii as ripening flora in controlled production of soft smear cheese from reconstituted milk: protein degradation.

Model smear soft cheeses, prepared with Debaryomyces hansenii and Brevibacterium linens as ripening starters, were ripened under aseptic conditions. Results of the cheese-making trials, in triplicate, were similar and showed similar patterns of protein degradation. In all of the trials, the acid-soluble nitrogen and nonprotein nitrogen (NPN) indexes and NH3 concentrations of the rind were low until d 10. The acid-soluble nitrogen and NPN of the rind then increased to 100 and 18% of total nitrogen, respectively, at d 76. The NH3 concentrations remained low until d 24 and increased until d 70, reaching about 1.8 g of NH3/kg of DM, and then remained constant. The acid-soluble nitrogen and NPN indexes and NH3 concentrations in the inner cheese mass were lower than in the rind. They showed the same evolution, reaching about 18% for acid-soluble nitrogen, 10% for NPN, and 1.5 g of NH3/kg of DM. It was shown that the inner cheese pH and populations of D. hansenii and B. linens have an effect on proteolysis. Viable cell counts of D. hansenii and B. linens were correlated with the environmental conditions and with proteolytic products. The determining role of carbon source and NH3 diffusions on the cheese ripening process were confirmed.

Ammonia↗

An improved method for screening alpha-acetolactate producing mutants.

Alpha-Acetolactate-deficient Lactococcus lactis ssp. lactis biovar. diacelylactis are utilised in several industrial processes for producing diacetyl and alpha-acetolactate. They can be selected by screening after random mutagenesis. We improved a previously described screening method [Monnet, C., Schmitt, P., Diviès, C., 1997. Appl. Environ. Microbiol. 63, 793-795], which makes it possible to screen up to 1000 colonies per agar plate, whereas the previous method allowed to screen only 60 colonies per agar plate. The new screening method facilitates selection of alpha-acetolactate-deficient mutants.

Filtration↗

Effect of alpha-acetolactate decarboxylase inactivation on alpha-acetolactate and diacetyl production by Lactococcus lactis subsp. lactis biovar diacetylactis.

Strains of Lactococcus lactis subsp. lactis biovar diacetylactis deficient in alpha-acetolactate decarboxylase produce alpha-acetolactate. This unstable compound is a precursor of acetoin and an aromatic compound, diacetyl. Following random mutagenesis of strain CNRZ 483, alpha-acetolactate decarboxylase-negative mutant 483 M1 was selected. When grown in milk, its growth and acidification characteristics were similar to those of the parental strain. In anaerobic conditions, the parental strain produced 2.10 mM acetoin and less than 0.05 mM diacetyl. The mutant accumulated up to 2.11 mM alpha-acetolactate, which spontaneously degraded to acetoin and diacetyl. After 24 h of culture, the alpha-acetolactate concentration was only 0.49 mM and the acetoin and diacetyl concentrations reached 1.50 mM and 0.26 mM, respectively. Diacetyl production by both strains increased in aerobic conditions, as well as when citrate was added. In contrast to cultures of the parental strain, however, diacetyl and acetoin concentrations in mutant cultures continued to increase without reaching a plateau. The results also showed that diacetyl production by wild type L. lactis subsp. lactis biovar diacetylactis strains cannot be explained uniquely by the spontaneous decarboxylation of the alpha-acetolactate produced in the culture medium.

Journal Article↗

Effect of citrate on growth of Lactococcus lactis subsp. lactis in milk.

The effect of citrate on the growth of Lactococcus lactis subsp. lactis var. diacetylactis in milk has been investigated. Five strains of Lactococcus lactis subsp. lactis var. diacetylactis were compared to their citrate-negative variants, which lack the plasmid coding for citrate permease. In most cases, acidification kinetics and the final bacterial concentration of pure cultures of parental and variant strains did not differ significantly. Co-cultures of parental and variant strains, however, systematically tended towards the predominance of parental strains. Citrate metabolism is responsible for this change, since the predominance of citrate-positive strains was not observed in the absence of citrate. Continuous culture in milk enabled the difference in growth rates between the parental strain Lactococcus lactis subsp. lactis var. diacetylactis CDI1 and its citrate-negative variant to be quantified by following changes in the populations of the two co-cultured strains. At 26 degrees C, the growth rate of the parental strain was 7% higher than that of its citrate-negative variant. These results show that citrate metabolism slightly stimulates the growth of lactococci in milk.

Animals↗

Microbial dynamics of co- and separately entrapped mixed cultures of mesophilic lactic acid bacteria during the continuous prefermentation of milk.

Four strains of mesophilic lactic acid bacteria were separately or coentrapped in kappa-carrageenan/locust bean gum gel beads and used for continuous prefermentation of UHT skim milk in a stirred-tank bioreactor. Lactic acid and cell productivities of the immobilized cell bioreactor were particularly high and remarkably stable during eight weeks of continuous milk prefermentation (about 18 g h-1 l-1 of lactic acid and 4.9 x 10(12) CFU h-1 l-1, respectively, but important variations of the bacterial populations is prefermented milk and gel beads occurred in any case (co-or separate entrapment). The strain Lactococcus lactis subsp. lactis biovar diacetylactis CDII became dominant, accounting for approx. 90% (released cells) and 78% (immobilized cells) of the total population. Microscopic observations of sections of gels beads showed a progressive destructing of the bead surface with rupture and release of entrapped viable cells from peripheral cavities of the gel. It is believed that these cavities close again after releasing all or part of their cell content, entrapping the different strains of the mixed culture and initiating a new colonization step and a cross-contamination of the beads. On the other hand, experimentations over seven-week periods with pasteurized milk showed the high resistance of the immobilized cell bioreactor to psychrotrophic contamination.

Animals↗

Use of an immobilized cell bioreactor for the continuous inoculation of milk in fresh cheese manufacturing.

A system was developed to continuously acidify and inoculate skim milk for the production of fresh cheese. Four strains of mesophilic lactic acid bacteria were entrapped separately in kappa-carrageenan/locust bean gum gel beads and used in a stirred bioreactor operated at 26 degrees C with a 25% (v/v) gel load. The pH in the reactor was controlled at 6.0 by adding fresh milk using proportional integrated derived regulation. The bioreactor was operated during 8-h daily cycles for up to 7 weeks with different milks (heat treatment, dry matter content) and differing starting procedures. The heat treatment of the milk was an important factor for process performance: a dilution rate increase of 57% and an inoculation level decrease of 63% were observed with sterilized UHT skim milk (142 degrees C - 7.5 s) compared with pasteurized skim milk (72 degrees C - 15 s). The dry matter content of the milk (8-13% w/w) had no detectable effect on these parameters. A convenient starting procedure of the system was tested; steady-state was reached in less than 40 min following an interruption period of 16-60 h. These results combined with our published data on process performance show the feasibility of using an integrated immobilized cell bioreactor for milk prefermentation in cheese manufacture.

Animals↗

Isolation and properties of Lactococcus lactis subsp. lactis biovar diacetylactis CNRZ 483 mutants producing diacetyl and acetoin from glucose.

Following treatment with the mutagen N-methyl-N'-nitro-N-nitrosoguanidine, three mutants of Lactococcus lactis subsp. lactis biovar diacetylactis CNRZ 483 that produced diacetyl and acetoin from glucose were isolated. The lactate dehydrogenase activity of these mutants was strongly attenuated, and the mutants produced less lactate than the parental strain. The kinetic properties of lactate dehydrogenase of strain CNRZ 483 and the mutants revealed differences in the affinity of the enzyme for pyruvate, NADH, and fructose-1,6-diphosphate. When cultured aerobically, strain CNRZ 483 transformed 2.3% of glucose to acetoin and produced no diacetyl or 2,3-butanediol. Under the same conditions, mutants 483L1, 483L2, and 483L3 transformed 42.0, 78.9, and 75.8%, respectively, of glucose to C4 compounds (diacetyl, acetoin, and 2,3-butanediol). Anaerobically, strain CNRZ 483 produced no C4 compounds, while mutants 483L1, 483L2, and 483L3 transformed 2.0, 37.0, and 25.8% of glucose to acetoin and 2,3-butanediol. In contrast to the parental strain, the NADH balance showed that the mutants regenerated most of the NAD via NADH oxidase under aerobic conditions and by ethanol production under anaerobic conditions.

Acetoin↗

Influence of dilution rate and cell immobilization on plasmid stability during continuous cultures of recombinant strains of Lactococcus lactis subsp. lactis.

The influence of dilution rate and cell immobilization on plasmid stability in recombinant strains of Lactococcus lactis subsp. lactis was investigated during continuous cultures. The studied strains, L. lactis IL2682 and IL2683, contained plasmids pIL9 (Lac+), pIL205 (CmR) and plasmids pIL252 (low copy number) and pIL253 (high copy number), respectively, that conferred resistance to erythromycin. Plasmid pIL205 was remarkably stable. Dilution rate did not affect the rate of loss of plasmids pIL252 and pIL253 significantly. Nevertheless, the loss of plasmid pIL253 was apparent after a further 21 generations when the dilution rate was decreased from 0.70 h-1 to 0.55 h-1. Cell immobilization in beads of kappa-carrageenan/locust bean gum improved plasmid stability by factors of 4.5 for pIL253 and 6.5 for pIL252. Thus, 10% of cells containing plasmids pIL252 or pIL253 were still present after 370 or 540 generations, respectively, compared with 50 or 210 generations in free cell cultures.

Culture Media↗

Effect of Initial Oxygen Concentration on Diacetyl and Acetoin Production by Lactococcus lactis subsp. lactis biovar diacetylactis.

The production of aroma compounds (acetoin and diacetyl) in fresh unripened cheese by Lactococcus lactis subsp. lactis biovar diacetylactis CNRZ 483 was studied at 30 degrees C at different initial oxygen concentrations (0, 21, 50, and 100% of the medium saturation by oxygen). Regardless of the initial O(2) concentration, maximal production of these compounds was reached only after all the citrate was consumed. Diacetyl and acetoin production was 0.01 and 2.4 mM, respectively, at 0% oxygen. Maximum acetoin concentration reached 5.4 mM at 100% oxygen. Diacetyl production was increased by factors of 2, 6, and 18 at initial oxygen concentrations of 21, 50, and 100%, respectively. The diacetyl/acetoin concentration ratio increased linearly with initial oxygen concentration: it was eight times higher at 100% (3.3%) than at 0% oxygen (0.4%). The effect of oxygen on diacetyl and acetoin production was also shown with other lactococci. At 0% oxygen, specific activity of alpha-acetolactate synthetase (0.15 U/mg) and NADH oxidase (0.04 U/mg) was 3.6 and 5.4 times lower, respectively, than at 100% oxygen. The increasing alpha-acetolactate synthetase activity in the presence of oxygen would explain the higher production of diacetyl and acetoin. The NADH oxidase activity would replace the role of the lactate dehydrogenase, diacetyl reductase, and acetoin reductase in the reoxidation of NADH, allowing accumulation of these two aroma compounds.

Journal Article↗

Batch cultures of recombinant Lactococcus lactis subsp. lactis in a stirred fermentor. I. Effect of plasmid content on bacterial growth and on genetic stability in pure cultures.

The effect of plasmid introduction into Lactococcus lactis subsp. lactis IL2661 on the growth of this strain and on plasmid stability was studied in pure batch cultures. The plasmids used (coding for erythromycin or chloramphenicol resistance) were the following: pIL205 (42 kb), pIL252 (4.6 kb, 6-9 copies), pIL253 (4.8 kb, 45-85 copies) and pE194 (inserted in the chromosome). Growth and acidification of L. lactis subsp. lactis IL2661 were similar to those of the derived recombinant lactococci. The maximal population at the end of the fermentation (9 h) was about 1.1 +/- 0.3 x 10(10) cfu/ml, and maximal growth rate 0.92 +/- 0.07 h-1. Growth yield and lactic acid concentrations were 3.9 +/- 0.8 x 10(11) cfu/g lactose consumed and 25.6 +/- 2.3 g/l, respectively. Different levels of plasmid stability were detected. Plasmid pE194, and plasmids pIL252 and pIL253 in the absence of pIL205, were stable after 10 h of culture. A slight loss (1-2%) of pIL205 was observed in all strains. In the presence of pIL205, plasmids pIL252 and pIL253 were maintained in only 56-95% of the cells. This result suggested an incompatibility between pIL205 and pIL252 or pIL253.

Biotechnology↗

Batch cultures of recombinant Lactococcus lactis subsp. lactis in a stirred fermentor. II. Plasmid transfer in mixed cultures.

The transfer of plasmids was studied in a stirred fermentor in the course of mixed batch cultures combining recombinant strains of Lactococcus lactis subsp. lactis (donor strains) with L. lactis subsp. lactis CNRZ 268M3 (recipient strain). Donor strains contained one or two of the following plasmids (coding for erythromycin or chloramphenicol resistance): pIL205 (self-transmissible), pIL252, pIL253 (non-transmissible but mobilizable by pIL205, respectively small and large copy number) and pE194 (inserted in the chromosome). Only self-transmissible plasmid pIL205 was transferred, with frequencies ranging from 10(-7) to 10(-8) after 12 h of fermentation. These frequencies were 60-400 times lower than in unstirred M17 broth and 100,000 times lower than on agar medium. In the latter case, non-transmissible plasmids pIL252 and pIL253 were mobilized by pIL205 with a frequency of about 10(-5) - 10(-6).

Biotechnology↗

Freeze-drying of Streptococcus thermophilus: a comparison between the vacuum and the atmospheric method.

Frozen suspensions of Streptococcus thermophilus were freeze-dried in a vacuum or a fluidized adsorbent bed at atmospheric pressure. Optimum operating conditions for each process were defined. For the duration of processing and survival rate of bacteria, in each case vacuum freeze-drying seemed more satisfactory than atmospheric pressure freeze-drying. The use of reconstituted skimmed milk as a suspension medium provides good protection for S. thermophilus.

Atmospheric Pressure↗

Characterization of Peroxidase Secretion and Subcellular Organization of Phanerochaete chrysosporium INA-12 in the Presence of Various Soybean Phospholipid Fractions.

Stimulation of lignin peroxidase production by exogenous phospholipids depends on the composition of the phospholipid fraction prepared by using the Nattermann process. The fraction composed mainly of negatively charged phospholipids (NAT 89) was the most efficient source for exoprotein secretion by Phanerochaete chrysosporium INA-12. The results of biochemical marker assays and ultrastructural morphology determination by electron microscopy were correlated. Activities of succinate dehydrogenase, a mitochondrial marker, and cytochrome c oxidoreductase, an endoplasmic reticulum (ER) marker, were increased 1.3- and 2.2-fold, respectively, in the presence of NAT 89. Electron microscopy observations suggested that the amount of mitochondria and ER in culture containing phospholipids was increased at the optimum day of lignin peroxidase production. Therefore, phospholipids enhanced energetic metabolism of strain INA-12 and markedly modified fungus physiology. Since ER is involved in enzyme synthesis, we suggest that its increased amount in mycelium cultured with NAT 89 is directly associated with the higher production of lignin peroxidase.

Journal Article↗

Effect of Fermentation Conditions on Growth of Streptococcus cremoris AM2 and Leuconostoc lactis CNRZ 1091 in Pure and Mixed Cultures.

Two strains of mesophilic lactic acid bacteria, Streptococcus cremoris AM2 and Leuconostoc lactis CNRZ 1091, were grown in pure and mixed cultures in the presence or absence of citrate (15 mM) and at controlled (pH 6.5) or uncontrolled pH. Microbial cell densities at the end of growth, maximum growth rates, the pH decrease of the medium resulting from growth, and the corresponding acidification rates were determined to establish comparisons. The control of pH in pure cultures had no effect on L. lactis CNRZ 1091 populations. The final populations of S. cremoris AM2, however, were at least five times higher than when the pH was not controlled (4 x 10 vs. 2 x 10 CFU . ml). The pH had no effect on the growth rate of either strain. That of S. cremoris AM2 (0.8 h) was about twice that of L. lactis CNRZ 1091. When the pH fell below 5, the growth of both strains decreased or stopped altogether. Citrate had no effect on S. cremoris AM2, while final populations of L. lactis CNRZ 1091 were two to three times higher (3 x 10 CFU . ml); it had no effect on the maximum growth rates of the two strains. Citrate attenuated the pH decrease of the medium and reduced the maximum acidification rate of the culture by 50%, due to the growth of S. cremoris AM2. Acidification due to L. lactis CNRZ 1091, however, was very slight. Regardless of the conditions of pH and citrate, the total bacterial population in mixed culture was lower (by 39%) than that of the sum of each pure culture. Mixed culture improved the maximum growth rate of L. lactis CNRZ 1091 (0.6 h) by 50%, while that of S. cremoris AM2 was unaffected. The acidification rate of the growth medium in mixed culture, affected by the presence of citrate, resulted from the development and activity of S. cremoris AM2.

Journal Article↗

Control of Lignin Peroxidase Production by Phanerochaete chrysosporium INA-12 by Temperature Shifting.

There are two temperature optima connected with lignin peroxidase synthesis by Phanerochaete chrysosporium INA-12. One, at 37 degrees C, is for the mycelium-growing phase; the other, at 30 degrees C, is for the lignin peroxidase-producing phase. One of six extracellular proteins with ligninase activity increased when cultures were grown at 30 degrees C for the entire fermentation period or when cultures were grown at 37 degrees C for the first 2 days of incubation and then shifted to 30 degrees C, compared with the activity of control cultures grown at 37 degrees C for the entire fermentation period. The unsaturation of fatty acid (Delta/mole) of P. chrysosporium INA-12 mycelium decreased from 1.25 to 1.03 when the growth temperature was shifted from 20 to 40 degrees C.

Journal Article↗