Proliferation of Lactobacillus plantarum in solid-state fermentation of oats

In an attempt to introduce probiotic functionalities to breakfast cereals and similar food products, the technique of solid-state fermentation (SSF) was applied to cultivate Lactobacillus plantarum (NCIMB 8826) on oat bran and spent oats after lipid extraction by supercritical CO(2) extraction. When compared to the frequently favored submerged processes for bacterium incubation, SSF presents not only the potential of simple downstream processing but also a more natural growth environment for the target bacterium. Preliminary studies confirmed that oat bran contained balanced nutrients to support a 25-fold bacterium propagation within a range of moisture content from 50% to 58% after a 36-h cultivation. Limited hydrolysis of the raw materials by the enzyme complex from submerged incubation of Aspergillus awamori and A. oryzae to increase nutrient accessibility extended the exponential growth phase and enhanced bacterial growth by over 183-fold. The process with the most potential, however, was to simultaneously grow both fungi aerobically on the raw materials in solid state to achieve sufficient hydrolysis, followed by controlled fungal autolysis at 65 degrees C prior to anaerobic bacterium incubation. Following this process bacterium population reached a maximum of 7.3 x 10(9) cells in each gram of the fermented solids, corresponding to a 1653-fold increase from the point of inoculation.     

Enrichment of wheat bran byRhodotorula gracilis through solid-state fermentation

The potential oil-producing yeast Rhodotorula gracilis was found to produce higher yields of biomass (13.7 g/L) and lipids (20.3%) in a nitrogen-limited and economically cheaper medium (molasses without yeast extract) in a submerged fermentation system. But, when the yeast was grown on four different wheat bran media by solid-state fermentation technique, different media combinations affected the percent increase in biomass, protein, oil production, fatty acid profile and degree of saturation and unsaturation. The initial lipid content in the control medium was 3.5% while in a medium with wheat bran, molasses, and minerals it was 69.8%. The yeast did not produce alpha-amylase, amyloglucosidase and cellulolytic enzymes for the breakdown of wheat bran. The yeast produced red carotenoids, a precursor of vitamin B12 and some oligounsaturated fatty acids in the fermented product.     

Production of novel oligosaccharide oxidase by wheat bran solid-state fermentation

A search for oxidases that catalyze the oxidation of oligosaccharides has resulted in the isolation of several soil-derived fungus strains which produced novel oligosaccharide oxidases with different substrate specificity on wheat bran solid culture. One of these oxidases produced by Acremonium strictum T1 strain has been characterized. This enzyme showed high reactivity toward maltose, lactose, cellobiose and maltooligosaccharides composed of up to seven glucose units, and was named as glucooligosaccharide oxidase based on its substrate specificity. Strain T1 was subjected to a strain improvement program, and an enzyme hyper-producing mutant strain T1-38 was selected. This mutant strain produced glucooligosaccharide oxidase 75 times higher than the wild type strain T1. When cultivated in a solid medium comprised of 1 part of wheat bran and 1 part of water (w/w), enzyme activity reached a maximum level of 6 units per g of culture medium after 4 days cultivation. Characteristics of the enzyme including the substrate specificity were compared with two other novel oligosaccharide oxidases isolated in this laboratory. Batch type conversion of lactose to lactobionic acid using crude enzyme was also discussed.     

Malt sprout extract medium for cultivation of Lactobacillus plantarum protective cultures

AIMS: The aim was to develop a cheap cereal-based alternative medium for the large-scale production of biopreservative Lactobacillus plantarum VTT E-79098. We examined the effect of growth medium and pH control on the cell yield of Lact. plantarum E-79098 and the antimicrobial activity of the cell-free extracts. METHODS: Fermentations using a novel Malt Sprout Extract Medium (MSE) were performed with different pH regimes. The antimicrobial activity of the cell-free extracts against Pantoea agglomerans VTT E-90396 and Fusarium avenaceum VTT D-80147 was assessed with automated turbidometry. SIGNIFICANCE AND IMPACT OF THE STUDY: When compared with MRS, the MSE medium cultures produced equal growth yields of Lact. plantarum VTT E-79098 and enhanced antimicrobial potential against the Gram-negative bacterium P. agglomerans and a Fusarium fungus. The MSE medium can be used as a low-cost alternative to MRS for producing high cell yields and good antimicrobial activity of Lact. plantarum.     

Comparison of phytase production on wheat bran and oilcakes in solid-state fermentation by Mucor racemosus

Comparisons were made for phytase production using wheat bran (WB) and oilcakes as substrates in solid-state fermentation (SSF) by Mucor racemosus NRRL 1994. WB was also used as mixed substrate with oil cakes. Sesame oil cake (SOC) served as the best carbon source for phytase synthesis by the fungal strain as it gave the highest enzyme titres (30.6 U/gds). Groundnut oil cake (GOC) also produced a reasonably good quantity of enzyme (24.3 U/gds). Enzyme production on WB was surprisingly much less (almost 3.5 times less in comparison to SOC). Mixing WB with SOC (1:1 ratio) resulted in better phytase activity (32.2 U/gds). Optimization of various process parameters such as incubation time, initial moisture content and inoculum concentration was carried out using the single variable mode optimization technique. Under optimized conditions, the production of phytase reached 44.5 U/gds, which was almost 1.5-fold higher than the highest yield obtained with any individual substrate used in this study and was more than 4-fold higher than that obtained from WB.     

氮素形态对专用小麦中后期根际土壤微生物和酶活性的影响

采用盆栽方法研究了酰胺态氮、铵态氮和硝态氮对强筋小麦(Triticum aestivum L.)"豫麦34"、中筋小麦"豫麦49"和弱筋小麦"豫麦50"生育中后期根际微生物和土壤酶活性的影响.结果表明,专用小麦根际真菌、细菌、放线菌数量和土壤脲酶、蛋白酶、硝酸还原酶活性以及根际pH值对氮素形态的反应不同."豫麦34"施用硝态氮,对根际土壤真菌、细菌(除成熟期外)和放线菌数量均具有明显的促进作用;"豫麦49"施用铵态氮,根际土壤细菌和放线菌数量最大,根际真菌数量在孕穗期和开花期以酰胺态氮处理最大,而成熟期以硝态氮处理最大;"豫麦50"施用硝态氮,对根际土壤真菌、细菌和放线菌数量均具有明显的促进作用.不同专用小麦品种均表现为在酰胺态氮处理下,根际土壤脲酶活性最高;在铵态氮处理下,根际土壤蛋白酶活性最高;在硝态氮处理下,根际土壤硝酸还原酶活性和pH值最高.     

Fermentation of quinoa and wheat slurries by Lactobacillus plantarum CRL 778: proteolytic activity

Quinoa fermentation by lactic acid bacteria (LAB) is an interesting alternative to produce new bakery products with high nutritional value; furthermore, they are suitable for celiac patients because this pseudo-cereal contains no gluten. Growth and lactic acid production during slurry fermentations by Lactobacillus plantarum CRL 778 were greater in quinoa (9.8 log?cfu/mL, 23.1 g/L) than in wheat (8.9 log?cfu/mL, 13.9 g/L). Lactic fermentation indirectly stimulated flour protein hydrolysis by endogenous proteases of both slurries. However, quinoa protein hydrolysis was faster, reaching 40–100 % at 8 h of incubation, while wheat protein hydrolysis was only 0–20 %. In addition, higher amounts of peptides (24) and free amino acids (5 g/L) were determined in quinoa compared to wheat. Consequently, greater concentrations (approx. 2.6-fold) of the antifungal compounds (phenyllactic and hydroxyphenyllactic acids) were synthesized from Phe and Tyr in quinoa by L. plantarum CRL 778, an antifungal strain. These promising results suggest that this LAB strain could be used in the formulation of quinoa sourdough to obtain baked goods with improved nutritional quality and shelf life, suitable for celiac patients.     

Biobleaching of banana fibre pulp using <Emphasis Type="Italic">Bacillus subtilis</Emphasis> C O1 xylanase produced from wheat bran under solid-state cultivation

A cellulase-free xylanase produced by Bacillus subtilis C 01 from wheat bran under solid-state cultivation was tested for its efficacy in biobleaching of raw banana fibre and banana pulp obtained through a mechanical pulping process. Banana pulp samples treated with crude xylanase (450 nkat g⁻¹ pulp) resulted in a 19.6% increase in the brightness as compared to untreated pulp. The presence of chromophores, hydrophobic compounds and an increased reducing sugar (10.79 mg g⁻¹ pulp) quantity in the bleached solution after enzymatic treatment indicated the removal of materials that were absorbed at 237 nm from the banana pulp.     

Significance of bed porosity, bran and specific surface area in solid-state cultivation of Aspergillus oryzae

In this paper, the effects of bed porosity, bran and specific surface area on the oxygen uptake rate and alpha-amylase production during growth of Aspergillus oryzae on wheat grain and wheat-flour substrate are reported. The high oxygen uptake rate found during cultivation of A. oryzae on wheat-flour substrate was not reached on wheat grain. This is mainly due to the bran of the wheat grain. Using wheat-flour substrates, it was shown that extra bed porosity increased the alpha-amylase production and oxygen uptake rates. Furthermore, the peak oxygen uptake rate decreased with increasing surface area-volume ratio of the substrate particles, while the alpha-amylase production and the cumulative oxygen uptake per gram of initial substrate dry matter increased. The present work does not support a direct correlation between aerial mycelia and enzyme production. There is, however, a correlation between the alpha-amylase yield and the cumulative oxygen uptake (not the uptake rate). This implies that aerial mycelia could accelerate alpha-amylase production even if they do not increase the yield.     

益生乳酸菌Lactobacillus casei Zhang和Lactobacillus plantarum P8对全价饲料pH及微生物类群变化的研究

研究了益生乳酸菌干酪乳杆菌Zhang(Lactobacillus casei Zhang)和植物乳杆菌P8(Lactobacillus planta-rum P8)对全价饲料pH及微生物类群变化的影响。分别将L.casei Zhang、L.plantarum P8单一菌种及复合菌种(11)以6.30 lg cfu/g的接种总量发酵全价饲料,测定25℃10 d发酵期间全价饲料pH和微生物类群的变化,应用选择培养基测定发酵饲料中的乳酸菌及杂菌(酵母菌、霉菌、大肠菌群、芽胞杆菌和梭状芽胞杆菌)的动态变化,应用RT-PCR技术测定试验组中的L.casei Zhang和L.plantarum P8的动态变化。结果显示,试验组pH下降显著,发酵10 d时,L.casei Zhang、L.plantarum P8单一菌种和复合菌种发酵饲料的pH分别为4.23、4.24和4.22,显著低于对照组(P0.05);L.casei Zhang、L.plantarum P8单一菌种和复合菌种发酵饲料中的L.casei Zhang、L.plantarum P8活菌数分别为8.91、8.89、6.58和8.69 lg cfu/g。发酵期间,试验组中酵母菌、霉菌、大肠菌群、芽胞杆菌及梭状芽胞杆菌活菌数显著低于对照组(P0.05),其中L.plantarum P8单一菌种发酵和复合菌种发酵对杂菌抑制效果显著优于L.casei Zhang单一菌种发酵(P0.05)。结果表明,全价饲料经L.casei Zhang、L.plantarum P8发酵可以显著降低其pH,抑制其中杂菌的生长,同时L.casei Zhang、L.plantarum P8在饲料中具有良好的稳定性。     

Properties of the Probiotic Strain Lactobacillus plantarum 8-RA-3 Grown in a Biofilm by Solid Substrate Cultivation Method

The biofilm formation took place in 48?h within the solid substrate cultivation of Lactobacillus plantarum 8-RA-3 strain on the wheat bran saturated with the MRS medium. The drying of the bran fermented by lactobacilli resulted in a decrease in the number of colony-forming units (CFU) from 23.0?×?10⁸ to 6.9?×?10⁵?CFU/g in daily samples and to less than 10⁴?CFU/g in 2- and 3-day samples. However, according to the fluorescence-based live/dead assay data, more than 40?% of the non-cultured bacteria were viable. As a result of mice kept on a diet with the introduction of bran fermented by Lact. plantarum 8-RA-3 for 72?h into the fodder, a recovery of normal level of intestinal lactobacilli, inhibited by administration of antibiotic was noted. The strain genetically identical to the Lact. plantarum 8-RA-3 was isolated from the feces of these mice. The results indicate that solid substrate cultivated Lact. plantarum 8-RA-3 strain formed a biofilm. Once dried and transferred into a non-cultured state, biofilm cells retained its viability and biological activity.     

1株植物乳杆菌生物学特性的研究

研究1株植物乳杆菌(N3)的生物学特性,实验结果表明该菌能耐受80~85℃的高温和0.20kg/cm2蒸汽压力;直接分解玉米淀粉的乳酸产率为7.05%(36 h)和8.19%(48 h);能耐受pH为4.5的酸性环境;在人工胃液中的活菌数为4.1×106CFU/g;对金霉素、土霉素、痢特灵和氟哌酸等抗生素的敏感性强,而对防霉剂和脱霉素不敏感。植物乳杆菌(N3)是1株优良的益生素生产菌。     

Effect of nitrogen source on lignocellulolytic enzyme production by white-rot basidiomycetes under solid-state cultivation

Summary The effect of additional nitrogen sources on lignocellulolytic enzyme production by four species of white-rot fungi (Funalia trogii IBB 146, Lentinus edodes IBB 363, Pleurotus dryinus IBB 903, and P. tuberregium IBB 624) in solid-state fermentation (SSF) of wheat straw and beech tree leaves was strain- and substrate-dependent. In general, the yields of hydrolytic enzymes and laccase increased by supplementation of medium with an additional nitrogen source. This stimulating effect of additional nitrogen on enzyme accumulation was due to higher biomass production. Only xylanase specific activity of P. dryinus IBB 903 and laccase specific activity of L. edodes IBB 363 increased significantly (by 66% and 73%, respectively) in SSF of wheat straw by addition of nitrogen source to the control medium. Additional nitrogen (20 mM) repressed manganese peroxidase (MnP) production by all fungi tested. The study of the nitrogen concentration effect revealed that 10 mM peptone concentration was optimal for cellulase and xylanase accumulation by P. dryinus IBB 903. While variation of the peptone concentration did not cause the change in MnP yield, elevated concentrations of this nutrient (20–40 mM) led to a 2–3-fold increase of P. dryinus IBB 903 laccase activity. About 10–20 mM concentration of NH₄NO₃ was optimal for cellulase and xylanase production by F. trogii IBB 146. However, neither the laccase nor the MnP yield was significantly changed by the additional nitrogen source.     

Genome sequence of the naturally plasmid-free Lactobacillus plantarum strain NC8 (CCUG 61730)

Lactobacillus plantarum is a highly versatile lactic acid bacterium found in various ecological niches, such as fermented vegetable, meat, and dairy products and the gastrointestinal tract. We sequenced the genome of L. plantarum NC8, a naturally plasmid-free strain, which has been used as a model strain in many laboratories worldwide.     

Production of cellulases and hemicellulases by Penicillium echinulatum grown on pretreated sugar cane bagasse and wheat bran in solid-state fermentation

AIM: To evaluate the solid-state fermentation (SSF) production of cellulase and hemicellulases (xylanases), by Penicillium echinulatum 9A02S1, in experiments carried out with different concentrations of the pretreated sugar cane bagasse (PSCB) and wheat bran (WB). METHODS AND RESULTS: This study reports the production of xylanolytic and cellulolytic enzymes by P. echinulatum 9A02S1 using a cheap medium containing PSCB and WB under SSF. The highest amounts of filter paper activity (FPA) could be measured on mixtures of PSCB and WB (32.89 +/- 1.90 U gdm(-1)). The highest beta-glucosidase activity was 58.95 +/- 2.58 U gdm(-1) on the fourth day. The highest activity for endoglucanases was 282.36 +/- 1.23 U gdm(-1) on the fourth day, and for xylanases the activity was around 10 U gdm(-1) from the second to the fourth day. CONCLUSIONS: The present work has established the potential of P. echinulatum for FPA, endoglucanase, beta-glucosidase and xylanase productions in SSF, indicating that WB may be partially substituted by PSCB. SIGNIFICANCE AND IMPACT OF THE STUDY: The incorporation of cheap sources, such as sugar cane bagasse, into media for the production of lignocellulose enzymes should help decrease the production costs of enzymatic complexes that can hydrolyse lignocellulose residues for the formation of fermented syrups, thus contributing to the economic production of bioethanol.     

The effect of temperature and Lactobacillus amylovorus and Lact. plantarum, applied at ensiling, on wheat silage

The effects of applying Lactobacillus plantarum and Lact. amylovorus at ensiling on wheat silage stored at 25 and41 °C was studied under laboratory conditions. The inoculants were applied at 10⁶ cfu g⁻¹.Silages with no additives served as controls. Three jars per treatment were sampled on days 2, 8 and 60 after ensiling, for chemical and microbiological analyses. After the ensiling period, the silages were subjected to an aerobic stability test. The control and Lact. plantarum inoculated wheat fermented faster at 25 than at 41 °C, whereas silages inoculated with Lact. amylovorus fermented faster at 41 °C. This was apparent from the rate of pH decrease and from the contents of residual sugars and lactic acid in the final silages. The numbers of lactobacilli in the control and Lact. plantarum silages at 41 °C after 2 and 8 days of ensiling were lower than in the corresponding silages at 25 °C. For the Lact. amylovorus silage the opposite held true. The control silages at both temperatures and the Lact. plantarum silage at 41 °C were the most stable silages under aerobic exposure.     

Purification and genetic characterization of plantaricin NC8, a novel coculture-inducible two-peptide bacteriocin from Lactobacillus plantarum NC8

A new, coculture-inducible two-peptide bacteriocin named plantaricin NC8 (PLNC8) was isolated from Lactobacillus plantarum NC8 cultures which had been induced with Lactococcus lactis MG1363 or Pediococcus pentosaceus FBB63. This bacteriocin consists of two distinct peptides, named alpha and beta, which were separated by C(2)-C(18) reverse-phase chromatography and whose complementary action is necessary for full plantaricin NC8 activity. N-terminal sequencing of both purified peptides showed 28 and 34 amino acids residues for PLNC8 alpha and PLNC8 beta, respectively, which showed no sequence similarity to other known bacteriocins. Mass spectrometry analysis showed molecular masses of 3,587 Da (alpha) and 4,000 Da (beta). The corresponding genes, designated plNC8A and plNC8B, were sequenced, and their nucleotide sequences revealed that both peptides are produced as bacteriocin precursors of 47 and 55 amino acids, respectively, which include N-terminal leader sequences of the double-glycine type. The mature alpha and beta peptides contain 29 and 34 amino acids, respectively. An open reading frame, orfC, which encodes a putative immunity protein was found downstream of plNC8B and overlapping plNC8A. Upstream of the putative -35 region of plNC8B, two direct repeats of 9 bp were identified, which agrees with the consensus sequence and structure of promoters of class II bacteriocin operons whose expression is dependent on an autoinduction mechanism.