Heterofermentative Strains Research Articles

220 Effect of silage inoculants containing different combinations of homo- and heterofermentative strains on fermentation, aerobic stability, and hydrocyanic acid content of sorghum whole-plant silage

220 Effect of silage inoculants containing different combinations of homo- and heterofermentative strains on fermentation, aerobic stability, and hydrocyanic acid content of sorghum whole-plant silage

Using pre-fermented sugar beet pulp as a growth medium to produce Pleurotus ostreatus mycelium for meat alternatives

This study aimed to determine the compatibility of pre-fermented sugar beet pulp to support the growth of Pleurotus ostreatus mycelium in submerged fermentation. The goal was to create a meat alternative based on mycelial-fermented pulp. It was further explored whether pre-fermentation with lactic acid bacteria (LAB) on the pulp increased meat-like properties, such as aroma, springiness, and hardness, in the final product. Three strains were selected from a high throughput screening of 105 plant-derived LAB based on their acidification and metabolite production in the pulp. Two homofermentative strains (Lactococcus lactis) and one heterofermentative strain (Levilactobacillus brevis) were selected based on their low ethanol production, high lactic acid production, and overall acidification of the pulp. Mycelium of P. ostreatus was grown in submerged fermentations on the pre-fermented pulp, and the biomass was removed by centrifugation. The fungal strain consumed all available sugars and acids and released arabinose to the media. Volatiles were detected using GC–MS, and a large increase in concentrations of hexanal, 1-octen-3-ol, and 2-octenal was measured. Concentration of 1-octen-3-ol was lower in the pre-fermented samples vs. the non-pre-fermented. LC-MS amino acid analysis showed the presence of all essential amino acids on day 0 and 7 of fermentation. The highest concentration of amino acids was for glutamic acid/glutamine and aspartic acid/asparagine. A decrease in all amino acids after 7 days of fungal fermentation was measured for all fermentations. The decrease was more significant for pre-fermented samples. This was also confirmed through a total protein determination, except for samples pre-fermented with Lactococcus lactis strain NFICC142 which increased in total protein content after fungal fermentation. The protein digestibility increased after fungal fermentation, and the highest increase was seen for non-pre-fermented samples. The springiness of the fermented product indicated similarities to meat alternatives, while the hardness was much lower than other meat alternatives. The results indicate that dried sugar beet pulp can be used for submerged cultivation of P. ostreatus, but that pre-fermentation does not improve the physical or nutritional properties of the end product significantly, except for an increased protein content for NFICC142 pre-fermented media. This is the first known attempt to use LAB and P. ostreatus in mixed fermentation to produce fungal mycelium, as well as the first attempt at using SBP in a liquid fermentation for mycelial production of P. ostreatus.

Comparison of the Chemical and Microbial Composition and Aerobic Stability of High-Moisture Barley Grain Ensiled with Either Chemical or Viable Lactic Acid Bacteria Application

This experiment was aimed at comparing how a chemical additive or an inoculant would affect the dry matter (DM) losses, fermentation pattern, yeast and mold counts, and aerobic stability (AS) of the ensiled high-moisture barley grain (675 g kg−1 DM). Crimped barley grain was ensiled with or without chemical additive AIV Ässä Na and an inoculant SiloSolve FC, totaling three treatments (1 × 3 factorial scheme) for the fermentation periods lasting 7, 14, 28, 60, and 90 days. The application of a chemical additive showed higher pH levels, retained water-soluble carbohydrates (WSCs) better, lowered DM loss, and reduced concentrations of fermentation products. Barley grain treated with chemicals initially showed a marked reduction in yeast and fungal growth and a higher AS up to day 60 of storage but became less effective in later stages of storage, leading to decreased AS. Barley grain inoculated with homo- and heterofermentative strains had decreased silage pH; its WSC content was similar to control and had higher content of weak acids and 1,2 propanediol, reducing mold and yeast counts. With fermentation duration from 28 days and beyond (i.e., 90 days), the inoculant treatment achieved the longest AS and the lowest increase in pH and weight loss during the period of air exposure.

Development of gluten-free bread biotechnology using fermented scald

The article is devoted to the development of gluten-free bread technology with scalded flour, fermented by lactic acid bacteria and yeast isolated previously from samples of good quality gluten-free sourdough. The species affiliation of new strains of lactic acid bacteria and yeast isolated from samples of good quality gluten-free starter cultures was established. The antagonistic and acid-forming activity of lactic acid bacteria and the fermentation activity of yeasts were investigated. An increase in the content of volatile acids in fermented scald and the acidity of finished products was established when using scald fermented with a heterofermentative strain L.brevis E139. As a result of the research, a biotechnology of fermented scald was developed, which makes it possible to obtain gluten-free bread with increased physical, chemical, organoleptic quality indicators and safety for diet therapy for celiac disease. The influence of the sourdough with a new microbial composition on the physico-chemical, organoleptic quality indicators of finished products and their resistance to mold and potato disease was studied.

Inoculation with heterofermentative strains <i>Lentilactobacillus buchneri</i> CNCM 40788 and <i>Lentilactobacillus hilgardii</i> CNCM I-4785 either alone or combined improves fermentation and aerobic stability of ensiled triticale (X-triticosecale)

<abstract> <p>Triticale, a small-grain forage, was harvested for an ensiling experiment evaluating different silage inoculants. Fresh material (mean dry matter 404.1 g kg<sup>−1</sup>) was wilted, chopped, and assigned to one of four treatment groups including water only (Control), heterofermentative strains <italic>Lentilactobacillus buchneri</italic> (LB), <italic>Lentilactobacillus hilgardii</italic> (LH), or combination (Combo) of both. Inoculants were applied at 4.0 x 10<sup>5</sup> CFU per g of fresh forage, and the Combo contained both strains at 2.0 x 10<sup>5</sup> CFU per g. Treated forage was packed into 7.57 L mini-silos for openings after 15, 30, and 130 d. Samples were collected at each opening for microbial enumeration of lactic acid bacteria, enterobacteria, yeasts, and moulds. Additional samples were collected for analysis of fermentation profiles, and nutritional analysis following dry matter determination and grinding. Aerobic stability was also evaluated at each opening through a 10-d period. Fermentation profiles were notably affected, including increases in acetic acid (g/kg DM) in LB and Combo treated silage after 15 d which resulted in reduced pH. Aerobic stability was vastly improved through inoculation by prolonging the time to reach 2˚ C above ambient, which was likely a result of decreased yeast counts. Our research validates the use of combined heterofermentative lactic acid bacteria strains on an ensiled small-grain specie, and further agrees with previous findings of prolonged aerobic stability through increases in lactate and acetate in response to co-inoculation.</p> </abstract>

Preservation characteristics and bacterial communities of crimped ensiled barley grains modulated by moisture content and additive application.

Information about the relationships between preservation characteristics and main bacterial communities of fermented feeds can guide decision making during feed preservation and silage additive development. The objective was to evaluate fermentation quality, aerobic stability, microbial quality and bacterial profile of crimped barley grains ensiled under three moisture contents (MC): 228 (low MC), 287 (medium MC) and 345 (high MC) g/kg fresh matter; and using four additive treatments: 1. Control (CONT), 2. Formic and propionic acid-based additive (FPA), 3. Inoculation with homofermentative and heterofermentative strains of lactic acid bacteria (LAB), and 4. Salt-based additive (SALT). There was a quadratic effect (p < 0.05) of incremental MC on pH where greater decline happened from low (5.81) to medium (4.83) MC than from medium to high (4.28) MC, while lactic acid concentration and aerobic stability increased in a linear manner (p < 0.05). Ammonia-N and acetic acid concentrations increased quadratically (p < 0.05) with increasing levels of MC. The effects of additives depended on MC so that improvements in preservation characteristics in response to LAB and SALT were observed at medium and high MC, while FPA was effective at all MC levels. A minor shift was observed in bacterial ecology from raw material towards low MC samples, with Erwiniaceae sp., Enterobacterales spp. and Pseudomonas dominating the fermentation. A major change occurred in medium and high MC materials, where Fructilactobacillus dominated the fermentation in CONT, FPA and SALT silages. LAB-treated silages at medium and high MC resulted in a distinguished pattern with dominance of Lentilactobacillus followed by Lactiplantibacillus. Most abundant communities in the samples, such as Fructilactobacillus, Erwiniaceae sp., Enterobacterales spp. and Pseudomonas, were correlated with several fermentation characteristics. Our results showed that crimped barley grains could be successfully ensiled under various MC and additive treatments. Low MC feeds had higher risk to be aerobically unstable while high MC resulted in more extensive fermentation, with potentially poor fermentation quality. The suitable additive depends on the raw material characteristics as LAB and SALT require relatively high MC to be effective, while FPA showed consistent improvements over all MC levels used in the current study. Awareness of the MC of grain prior to ensiling allows to identify the risks to preservation quality and provides information for choosing an effective additive.

Homo- and heterofermentative lactobacilli are distinctly affected by furanic compounds.

Second generation (2G) ethanol is produced using lignocellulosic biomass. However, the pre-treatment processes generate a variety of molecules (furanic compounds, phenolic compounds, and organic acids) that act as inhibitors of microbial metabolism, and thus, reduce the efficiency of the fermentation step in this process. In this context, the present study aimed to investigate the effect of furanic compounds on the physiology of lactic acid bacteria (LAB) strains that are potential contaminants in ethanol production. Homofermentative and heterofermentative strains of laboratory LAB, and isolated from first generation ethanol fermentation, were used. LAB strains were challenged to grow in the presence of furfural and 5-hydroxymethylfurfural (HMF). We determined that the effect of HMF and furfural on the growth rate of LAB is dependent on the metabolic type, and the growth kinetics in the presence of these compounds is enhanced for heterofermentative LAB, whereas they are inhibitory to homofermentative LAB. Sugar consumption and product formation were also enhanced in the presence of furanic compounds for heterofermentative LAB, who displayed effective depletion kinetics when compared to the homofermentative LAB. Homo- and heterofermentative LAB are affected differently by furanic compounds, in a way that the latter type is more resistant to the toxic effects of these inhibitors. This knowledge is important to understand the potential effects of bacterial contamination in 2G bioprocesses.

Anti-Salmonella activity of lactobacilli from different habitats

Lactic acid bacteria (LAB) may contribute to the food safety. In the present study, the antagonistic activity of 45 Bulgarian Lactobacillus homo- and heterofermentative strains of human and dairy origin, and 4 multibacterial formulas against Salmonella enterica subsp. enterica serovar Typhimurium (S. Typhimurium) was assessed. In vitro tests were performed in different model systems - milk, soymilk, De Man Rogose Sharp (MRS) medium, to simulate real conditions in the food chain. The highest antagonistic activity was observed with cell-free supernatants of exponential MRS broth cultures of the strains isolated from breast milk, followed by lactobacilli from white brined and green cheese. The detected antimicrobial activity against the pathogen was strain-specific and depended on the culture conditions. Lactobacillus (reclassified as Lactoplantibacillus) plantarum strains, cultivated in skimmed milk and whey protein medium, were able to inhibit S. Typhimurium growth, while a limited inhibitory activity was detected for fermented soymilks. A bacteriocinogenic Ligilactobacillus (the previous Lactobacillus) salivarius strain reduced the number of living pathogenic cells during co-cultivation in whole milk. The inhibition was significant only when L. salivarius was inoculated in predominance. In case of underrepresented LAB number, S. Typhimurium over-growth was observed. Eight lactobacilli in combination as a multibacterial co-culture expressed synergic antagonistic effect against Salmonella and were pre-selected as promising. Further characterisation of their active metabolites, however, is needed before their classification as bio-protective agents.

Biotechnology of Newly Created Bacterial Composition for Siloing Based on Lactic Acid Bacteria

Peculiarities of high-quality silage production are the use of biological products based on lactic acid bacteria. The composition of such starters varies greatly according to the use of bacterial cultures, so among the starters available on the market, the range of their effectiveness is also different. It is very common to use a one-sided approach to the choice of bacterial components, which in combination with imperfect production technology have low preservative activity. The study of combined preparations, which combine homo- and heterofermentative types of lactic acid fermentation, allows to stabilize the preservative properties throughout the ensiling time, and increase the aerobic stability of the silage after access of oxygen. Aim. Development of biotechnology of bacterial preparation for corn ensiling, optimization of cultivation conditions of newly created bacterial composition, and selection of cryoprotectants for its lyophilization. Methods. The combined preparation was created on the basis of heterofermentative strain Lactobacillus buchneri 3806 combining it in two- and three-strain compositions with other representatives of lactic acid bacteria, which are characterized by obligate homofermentative and facultative heterofermentative types of metabolism. Optimization of the environment and technological parameters was carried out using a central-compositional plan, further statistical analysis of the obtained data and determination of optimal values of input parameters according to the created mathematical model of optical density response. The effectiveness of the selected protective media was tested for the survival of bacteria after lyophilization. Results. The most effective bacterial composition was found during experiments: L. buchneri 3806, Enterococcus faecium C-8-12, L. plantarum 3216. The effectiveness of the obtained composition was tested by laboratory silage of corn. Tests of the drug based on the selected bacterial composition showed an improvement in the chemical composition of the silage compared to the untreated control and treated only with monoculture L. buchneri 3806, namely: there was a decrease in dry matter loss by 2.21% and 2.04%, 22 due to the increase of lactic acid content, and increase of aerobic stability of silage – 341 h against 57 h of the control sample, and 313 h in case of using monoculture. For the obtained bacterial composition, the culture medium of the following composition was optimized: base (hydrolyzed milk with the addition of the following components: monosubstituted potassium phosphate – 2 g/L; 5-aqueous manganese sulfate – 0.05 g/L; 7-aqueous magnesium sulfate – 0.2 g/L; twin-80 – 1.0 g/L); glucose – 19.7 g/L; yeast extract – 7.8 g/L; corn extract – 23.6 g/L; peptone – 9.1 g/L; sodium citrate – 6.6 g/L; sodium acetate – 3,4 g/L. Cultivation of the bacterial composition on an optimized medium made it possible to obtain the maximum biomass yield, at which the optical density was 2.01 units, which is almost twice as much as the value obtained by culturing the same composition in MRS medium. The optimal technological parameters of culturing the bacterial composition were established, namely the best growth was observed at a temperature of 36.4±0.4°C with constant maintenance of the pH value in the culture medium at the level of 6.5±0.1 units. In addition, the optimal composition of the protective medium containing sodium citrate, sucrose and agar was selected, and ensures the survival rate of lactic acid bacteria 98.4% after lyophilization. Conclusions. The newly formed bacterial composition can be used for the production of preparations for corn silage, and tested on other raw materials, in particular on some perennial legumes (alfalfa, clover), and the conditions of its production can be used to scale the technology.

New epiphytic strains of lactic acid bacteria improve the conservation of corn silage harvested at late maturity

We evaluated fifty-three strains of lactic acid bacteria (LAB) isolated from corn silages from different farms for use as inoculants in corn silages. Bacteria were characterized for growth and pH reduction in corn extract, growth at different temperatures and the ability to inhibit the growth of silage-spoilage microorganisms. The strains CCMA1362, 1363 and 1364 (Lactobacillus farraginis), CCMA1365 (L. plantarum), CCMA1366 (L. buchneri), and CCMA1367 (Pediococcus acidilactici) isolated from corn silage and CCMA170 strain (L. hilgardii) isolated from sugarcane showed good results in laboratory evaluation (good growth and reduction of pH in corn extract; good growth at different temperature and ability to inhibit silage-spoilage microorganisms). These strains were selected and evaluated in corn silages harvested at an advanced stage of physiological maturity (454 g/kg DM). Experimental silos were opened after 10, 32 and 100 days of storage. Crude protein, ash and starch contents were not affected by the inoculation or storage time. Inoculation with the obligate homofermentative strain CCMA 1367 (P. acidilactici) resulted in silages with the greatest DM loss and the lowest aerobic stability. Silages treated with the obligate heterofermentative strains (L. buchneri, L. hilgardii and L. farraginis) had the lowest DM loss, highest aerobic stability, good acid production and pH value, and lowest numbers of yeasts. The species L. farraginis is reported for the first time in studies with corn silage and in particular the CCMA1362 (L. farraginis) strain was effective in lowering DM loss and reducing the numbers of yeasts throughout the ensiling period. Our results are important because they show that silage conservation can be improved even in late maturity corn silage when treated with select lactic acid bacteria L. farraginis CCMA1362. More studies are needed to evaluate the efficiency of these strains under field conditions.

Potential of Lactic Acid Bacteria Isolated From Different Forages as Silage Inoculants for Improving Fermentation Quality and Aerobic Stability.

We aimed at isolating lactic acid bacteria (LAB) from different plant materials to study their crossed-fermentation capacity in silos and to find strains able to confer enhanced aerobic stability to silage. A total of 129 LAB isolates were obtained from lucerne (alfalfa), maize, sorghum, ryegrass, rice, barley, canola, Gatton panic, Melilotus albus, soy, white clover, wheat, sunflower, oat, and moha. Four Lactiplantibacillus plantarum subsp. plantarum strains (isolated from oat, lucerne, sorghum, or maize) were selected for their growth capacity. Identity (16S sequencing) and diversity (RAPD-PCR) were confirmed. Fermentative capacity (inoculated at 104, 105, 106, 107 CFU/g) was studied in maize silage and their cross-fermentation capacity was assessed in oat, lucerne, sorghum, and maize. Heterofermentative strains with the highest acetic acid production capacity conferred higher aerobic stability to maize silages. Regardless the source of isolation, L. plantarum strains, inoculated at a rate of 106 CFU/g, were effective to produce silage from different plant materials. From more than 100 isolates obtained, the application of a succession of experiments allowed us to narrow down the number of potential candidates of silage inoculants to two strains. Based on the studies made, L. plantarum LpM15 and Limosilactobacillus fermentum LfM1 showed potential to be used as inoculants, however further studies are needed to determine their performance when inoculated together. The former because it positively influenced different quality parameters in oat, lucerne, sorghum, and maize silage, and the latter because of its capacity to confer enhanced aerobic stability to maize silage. The rest of the strains constitute a valuable collection of autochthonous strains that will be further studied in the future for new applications in animal or human foods.

Technological Performance and Selection of Lactic Acid Bacteria Isolated from Argentinian Grains as Starters for Wheat Sourdough.

The selection of lactic acid bacteria strains is interesting for the development of sourdough to obtain wheat bread with improved technological and nutritional properties. In this work, the performance of ten different LAB strains isolated from diverse cereals in wheat sourdough was evaluated. Five facultative heterofermentative (Lactobacillus pentosus ES124, Lactobacillus paralimentarius ES259, Lactobacillus plantarum ES137, Lactobacillus plantarum ATCC8014 and Lactobacillus plantarum ES147), three obligately heterofermentative (Lactobacillus brevis ES253, Lactobacillus fermentum ES142 and Lactobacillus fermentum ES148) and two homofermentative (Pediococcus acidilactici ES22 and Enterococcus faecium ES74) lactic acid bacteria strains were evaluated in ten single strain fermentations. Sugar profile, volatile compounds and proteolytic ability of sourdoughs were analyzed. The microbiological counts showed that facultative heterofermentative strains presented higher counts than homofermentative sourdoughs. Monosaccharides (glucose and fructose), maltose and sucrose, isomaltose and dextrin were detected in fermentations with homofermentative strains whilst in those with heterofermentative strains maltose/sucrose, isomaltose and dextrin were found. L. paralimentarius ES259 and L. pentosus ES124 showed the highest diversity of volatile compounds. L. plantarum ES137 and P. acidilactici ES22 were the strains with the highest proteolytic activity. The technological performance allowed us to select LAB as starters to develop breads with specific rheological properties and final quality.

Application of mannitol producing Leuconostoc citreum TR116 to reduce sugar content of barley, oat and wheat malt-based worts

Achieving a high monosaccharide composition in malt wort is instrumental to achieve successful lactic acid bacteria fermentation of malt based beverages. The conversion of monosaccharides to alternative metabolites such as the sweet polyol, mannitol with heterofermentative strains presents a novel approach for sugar reduction and to compensate for the loss of sweetness. This work outlines the application of an adopted mashing regimen with the addition of exogenous enzymes to produce wort with high fructose content which can be applied to different malted grain types with consistently efficacious monosaccharide production for bacterial fermentation. The so produced worts are then fermented with Leuconostoc citreum TR116 a mannitol hyper-producer. Malted barley, oat and wheat were mashed to stimulate protein degradation and release of free amino acids along with the enzymatic conversion of starch to fermentable sugars. Amyloglucosidase and glucose isomerase treatment converted di- and oligo-saccharides to glucose and provided a moderate fructose concentration in malt worts which was consistent across the three cereals. Fructose was completely depleted during fermentation with Lc. Citreum TR116 and converted to mannitol with high efficiency (>90%) while overall sugar reduction was >25% in all malt worts. Differences in amino acid composition of malt worts did not significantly affect growth of Lc. Citreum TR116 but did affect the formation of the aroma compounds diacetyl and isoamyl alcohol. Organic acid production and acidification of wort was similar across cereal substrates and acetic acid formation was linked to yield of mannitol. The results suggest that differences in amino acid and fructose content of malt worts considerably change metabolite formation during fermentation with Lc. Citreum TR116, a mannitol hyper-producer. This work gives new insight into the development of consumer acceptable malt based beverages which will provide further options for the health conscious and diabetic consumer, an important step in the age of sugar overconsumption.

Endoglucanase improve the growth of homofermentative Lactobacillus spp. in ensilages

Endoglucanase, an important component of cellulases, is used as additives in ensiling of forage crops. However, its detailed role is unclear in ensilages. In the present study, two endoglucanases Cel5 and Cel9 produced by strain Paenibacillus panacisoli SDMCC050309, previously isolated from ensiled corn stover, were identified in the cultures by microcrystalline cellulose absorption coupled with zymogram analysis. After heterologously expressed in Escherichia coli DE3 and purified, these two proteins were biochemically characterized. Cel5 was 61 kDa and showed maximal activity at pH 7.0 and 45 °C, while the maximum activity was at pH 8.0 and 65 °C for Cel9 with 97 kDa in size. Both of them could degrade carboxymethyl cellulose into cellooligosaccharides, in which cellobiose and cellotriose could be used as substrates for the growth of homofermentative strains Lactobacillus plantarum CGMCC6888 and L. farciminis CCTCC AB2016237, but not for the heterofermentative strains L. brevis SDMCC050297 and L. parafarraginis SDMCC050300. Therefore, we concluded that the added endoglucanase contributed to enhance the growth of homofermentative lactic acid bacteria for high level of lactic acid production in ensilages.

Corrigendum to: Effect of Lactobacillus plantarum and Lactobacillus buchneri addition on fermentation, bacterial community and aerobic stability in lucerne silage

Changes in the microbial community are closely related to the fermentation of silage. However, how host genetic variation shapes the community structure of the silage microbiota and its metabolic phenotype is poorly understood. The objective of present study was to evaluate the effects of the application of the homo-fermentative Lactobacillus plantarum and hetero-fermentative Lactobacillus buchneri strains to lucerne silage on the fermentation characteristics, aerobic stability, and microbial community and their correlations. The three silages treated with L. plantarum or L. buchneri were well preserved and had significantly lower pH values, butyric acid, propionic acid, and ammonia-N concentrations, and significantly higher residual water-soluble carbohydrate, dry matter and lactic acid contents than the controls. The treated groups had more lactic acid bacteria and lower quantities of other bacteria in their microbial communities. Inoculation of lactic acid bacteria influenced the abundances of other bacteria and controlled the silage fermentation characteristics. L. buchneri inhibited the abundance of Enterobacter_ludwigii to increase the crude protein content, L. plantarum improve the neutral detergent fibre content by affecting the abundance of Arthrobacter_sp._Ens13. In conclusion, the application of L. plantarum and L. buchneri improved the quality of lucerne silage fermentation, and L. buchneri resulted in greater improvements after aerobic exposure.

Cobalamin production by Lactobacillus coryniformis: biochemical identification of the synthetized corrinoid and genomic analysis of the biosynthetic cluster.

BackgroundDespite the fact that most vitamins are present in a variety of foods, malnutrition, unbalanced diets or insufficient intake of foods are still the cause of vitamin deficiencies in humans in some countries. Vitamin B12 (Cobalamin) is a complex compound that is only naturally produced by bacteria and archea. It has been reported that certain strains belonging to lactic acid bacteria group are capable of synthesized water-soluble vitamins such as those included in the B-group, as vitamin B12. In this context, the goal of the present paper was to evaluate and characterize the production of vitamin B12 in Lactobacillus coryniformis CRL 1001, a heterofermentative strain isolated from silage.ResultsCell extract of L. coryniformis CRL 1001, isolated from silage, is able to correct the coenzyme B12 requirement of Salmonella enterica serovar Typhimurium AR 2680 in minimal medium. The chemical characterization of the corrinoid-like molecule isolated from CRL 1001 cell extract using HPLC and mass spectrometry is reported. The majority of the corrinoid produced by this strain has adenine like Coα-ligand instead 5,6-dimethylbenzimidazole. Genomic studies revealed the presence of the complete machinery of the anaerobic biosynthesis pathway of coenzyme B12. The detected genes encode all proteins for the corrin ring biosynthesis and for the binding of upper (β) and lower (α) ligands in one continuous stretch of the chromosome.ConclusionsThe results here described show for the first time that L. coryniformis subsp. coryniformis CRL 1001 is able to produce pseudocobalamin containing adenine instead of 5,6-dimethlbenzimidazole in the Coα-ligand. Genomic analysis allowed the identification and characterization of the complete de novo biosynthetic pathway of the corrinoid produced by the CRL 1001 strain.

Effect of heterofermentative lactic acid bacteria of DL-starters in initial ripening of semi-hard cheese

The role of heterofermentative lactic acid bacteria from dl-starters in ripening of semi-hard cheese was investigated using the strains Leuconostoc pseudomesenteroides PS12 and 1159, Leuconostoc mesenteroides subsp. cremoris T26 and Lactobacillus danicus 13M1. Control cheese was made with starter containing only homofermentative Lactococcus lactis subspecies. Leuc. mesenteroides subsp. cremoris T26 did not grow in cheese and started to decrease in number early, whereas the others grew and remained at a high number throughout the nine-week ripening period. None of the added heterofermentative strains affected proteolysis and total amount of amino acids; however, differences in the composition of amino acids were observed, and caused significant differences in the composition of volatile aroma compounds. Added strains increased the amount of secondary alcohols and mediated decreases in the amount of corresponding methyl ketones, diacetyl and acetoin. Eye formation was only affected by Lb. danicus through stimulation of late gas formation in cheeses.

Effects of additives on the fermentation and aerobic stability of grass silages and total mixed rations

AbstractMown herbage of timothy–meadow fescue (dry matter 218 (LDM) or 539 (HDM) g kg−1) was ensiled in laboratory silos to evaluate silage additives. For LDM silage, additives including formic acid (a blend of formic acid, sodium formate, propionic acid, benzoic acid, glycerol and another blend of formic acid and ammonium formate, both applied at 5 L t−1) were able to restrict fermentation and thereby improve intake potential of the silage. Aerobic stability (AS) of total mixed ration (TMR) was also improved. LDM grass treated with homofermentative lactic acid bacteria (hoLAB) resulted in silage containing lactic acid at 132 g kg−1 DM, ammonium‐N &lt;40 g kg−1 total N, and pH &lt; 3·8, and the AS was poor (&lt;36 h). The treatment including heterofermentative strain (Lactobacillus brevis) produced more acetic acid and better AS than hoLAB. Salt treatment (sodium benzoate, potassium sorbate, sodium nitrite) reduced pH compared to the Control treatment (3·89 vs. 4·24) and improved AS of TMR. The LDM Control silage had good AS, but the TMR based on it had poor AS. All additives were able to lower pH on HDM silages also, but other benefits of using additives were minimal. The treatment including L. brevis on HDM was able to improve AS of TMR.

Comparative genome analysis of the candidate functional starter culture strains Lactobacillus fermentum 222 and Lactobacillus plantarum 80 for controlled cocoa bean fermentation processes.

BackgroundLactobacillus fermentum 222 and Lactobacillus plantarum 80, isolates from a spontaneous Ghanaian cocoa bean fermentation process, proved to be interesting functional starter culture strains for cocoa bean fermentations. Lactobacillus fermentum 222 is a thermotolerant strain, able to dominate the fermentation process, thereby converting citrate and producing mannitol. Lactobacillus plantarum 80 is an acid-tolerant and facultative heterofermentative strain that is competitive during cocoa bean fermentation processes. In this study, whole-genome sequencing and comparative genome analysis was used to investigate the mechanisms of these strains to dominate the cocoa bean fermentation process.ResultsThrough functional annotation and analysis of the high-coverage contigs obtained through 454 pyrosequencing, plantaricin production was predicted for L. plantarum 80. For L. fermentum 222, genes encoding a complete arginine deiminase pathway were attributed. Further, in-depth functional analysis revealed the capacities of these strains associated with carbohydrate and amino acid metabolism, such as the ability to use alternative external electron acceptors, the presence of an extended pyruvate metabolism, and the occurrence of several amino acid conversion pathways. A comparative genome sequence analysis using publicly available genome sequences of strains of the species L. plantarum and L. fermentum revealed unique features of both strains studied. Indeed, L. fermentum 222 possessed genes encoding additional citrate transporters and enzymes involved in amino acid conversions, whereas L. plantarum 80 is the only member of this species that harboured a gene cluster involved in uptake and consumption of fructose and/or sorbose.ConclusionsIn-depth genome sequence analysis of the candidate functional starter culture strains L. fermentum 222 and L. plantarum 80 revealed their metabolic capacities, niche adaptations and functionalities that enable them to dominate the cocoa bean fermentation process. Further, these results offered insights into the cocoa bean fermentation ecosystem as a whole and will facilitate the selection of appropriate starter culture strains for controlled cocoa bean fermentation processes.Electronic supplementary materialThe online version of this article (doi:10.1186/s12864-015-1927-0) contains supplementary material, which is available to authorized users.

Complete genome sequence of Clostridium pasteurianum NRRL B-598, a non-type strain producing butanol

The strain Clostridium pasteurianum NRRL B-598 is non-type, oxygen tolerant, spore-forming, mesophilic and heterofermentative strain with high hydrogen production and ability of acetone–butanol fermentation (ethanol production being negligible). Here, we present the annotated complete genome sequence of this bacterium, replacing the previous draft genome assembly. The genome consisting of a single circular 6,186,879bp chromosome with no plasmid was determined using PacBio RSII and Roche 454 sequencing.