盐浓度对蚕豆酱发酵过程中原核微生物多样性及理化因子的影响

蚕豆酱是我国传统的发酵豆类调味品.盐作为一种重要的添加剂,对蚕豆酱的微生物群落结构和品质有着至关重要的影响.为探究不同盐浓度对蚕豆酱发酵过程中原核微生物和理化因子演替规律的影响.采用高通量测序技术对不同发酵时间蚕豆酱的原核微生物多样性进行了分析,并检测了其理化特征.结果表明,盐浓度不仅会显著影响pH、总酸、氨基酸态氮和...     

Enterotoxins and emetic toxins production by Bacillus cereus and other species of Bacillus isolated from Soumbala and Bikalga, African alkaline fermented food condiments

The ability of various species of Bacillus from fermented seeds of Parkia biglobosa known as African locust bean (Soumbala) and fermented seeds of Hibiscus sabdariffa (Bikalga) was investigated. The study included screening of the isolates by haemolysis on blood agar, detection of toxins in broth and during the fermentation of African locust bean using the Bacillus cereus Enterotoxin Reverse Passive Latex Agglutination test kit (BCET-RPLA) and the Bacillus Diarrhoeal Enterotoxin Visual Immunoassay (BDEVIA). Detection of genes encoding cytotoxin K (CytK), haemolysin BL (Hbl A, Hbl C, Hbl D), non-hemolytic enterotoxin (NheA, NheB, NheC) and EM1 specific of emetic toxin producers was also investigated using PCR with single pair and multiplex primers. Of 41 isolates, 29 Bacillus belonging to the species of B. cereus, Bacillus subtilis, Bacillus licheniformis and Bacillus pumilus showed haemolysis on blood agar. Using RPLA, enterotoxin production was detected for three isolates of B. cereus in broth and all B. cereus (9) in fermented seeds. Using BDEVIA, enterotoxin production was detected in broth as well as in fermented seeds for all B. cereus isolates. None of the isolates belonging to the other Bacillus species was able to produce enterotoxins either by RPLA or BDEVIA. Nhe genes were detected in all B. cereus while Hbl and CytK genes were detected respectively in five and six B. cereus strains. A weak presence of Hbl (A, D) and CytK genes was detected in two isolates of B. subtilis and one of B. licheniformis but results were inconsistent, especially for Hbl genes. The emetic specific gene fragment EM1 was not detected in any of the isolates studied.     

Processing and fermentation of the african locust bean (Parkia filicoidea Welw.) to produce dawadawa

African locust beans (Parkia filicoidea Welw.) were processed and fermented to the traditional West African condiment dawadawa. The beans were transformed to ‘processed substrate’ by boiling and dehulling, and then fermented at 37°C in static fermenter units. The overall mass balance and the mass balances of several bean components (moisture, fat, protein, ash and carbohydrate) were studied. It was found that 1.0 kg of raw beans (6.4% moisture) yielded 1.3 kg of processed substrate (63% moisture) which in turn was converted to 1.2 kg of dawadawa (65% moisture). The loss of bean solids during processing was due to the removal of adhering pulp and testa as well as to solids extraction during boiling and washing. Overall, 57% of the bean dry weight, 85% of the carbohydrate, 48% of the ash, 40% of the protein and 1% of the fat were lost in the conversion of raw beans to dawadawa. The dawadawa had considerably higher fat and protein contents than the raw beans and was a pleasant tasting food whereas the raw beans were inedible. Some microbiological aspects of the fermentation were also investigated. Normally prepared fermentation substrate was compared with both sterilised and radappertised beans in terms of conversion to dawadawa, number of colony forming units present, and the development of pH and titratable acidity. The presence of microorganisms was found to be obligatory for the conversion to proceed. Simultaneous increases in both pH and titratable acidity were observed during the fermentation.     

Genotyping of starter cultures of Bacillus subtilis and Bacillus pumilus for fermentation of African locust bean (Parkia biglobosa) to produce Soumbala

Bacillus spp. are the predominant microorganisms in fermented African locust bean called Soumbala in Burkina Faso. Ten strains selected as potential starter cultures were characterised by PCR amplification of the16S-23S rDNA intergenic transcribed spacer (ITS-PCR), restriction fragment length polymorphism of the ITS-PCR (ITS-PCR RFLP), pulsed field gel electrophoresis (PFGE) and sequencing of the 968-1401 region of the 16S rDNA. In previous studies, the isolates were identified by phenotyping as Bacillus subtilis and Bacillus pumilus. The phenotyping was repeated as a reference in the present study.The ITS-PCR and ITS-PCR RLFP allowed a typing at species level. The PFGE was more discriminative and allowed a typing at strain level. Full agreement with the phenotyping was observed in all cases. The sequencing of the 16S rDNA allowed the identification at species level with an identity from 97% to 100% comparing the sequences to those from the GenBank databases. The desired cultures of B. subtilis and B. pumilus from African locust bean fermentation were distinguished by ITS-PCR and ITS-PCR RLFP from Bacillus cereus and Bacillus sphaericus which sometimes occur in the beginning of the fermentation.     

Microbiological and chemical changes during the fermentation of African locust bean (Parkia biglobosa) to produce afitin, iru and sonru, three traditional condiments produced in Benin

African locust bean (Parkia biglobosa) was processed to produce afitin, iru and sonru, three different types of condiment from Benin. Whereas the fermentation of African locust bean to produce afitin is carried out for 24 h without using any additive, the fermentation for iru and sonru production takes place for 48 h with adding respectively "iku-iru" and "yanyanku" two traditional malvacene bean-based (Hibiscus sabdariffa) additives. The main microorganisms involved in the fermentation of the three condiments were Bacillus spp., although Staphylococcus spp. was found in lower number. The use of additive seems to enhance the initial Bacillus counts in iru and sonru, but also a slightly higher Staphylococcus count was observed. The number of staphylococci did not exceed 10(2) cfu/g and it was below the detection level at the end of the fermentation (24 h) in afitin; it reached a level of about 10(6) cfu/g at 12-18 h in iru and sonru followed by a decrease to about 5 x 10(4) cfu/g after 48 h of fermentation. The final pH and total basic volatile nitrogen content of iru and sonru are higher than those of afitin. Proximate composition changes are similar in both fermenting iru and sonru, but different in afitin, the latter showing a slight decrease of proteins of about 5% whereas for iru and sonru they increased to about 6%. Ash, crude fat and fibre contents also increased during the fermentation in the three products whereas total carbohydrates decreased significantly. Due to their long fermentation time, iru and sonru are very soft products as they are preferred by some consumers whereas afitin is preferred with cotyledons having less soft texture.     

Degradation of polysaccharides and non-digestible oligosaccharides by Bacillus subtilis and Bacillus pumilus isolated from Soumbala, a fermented African locust bean (Parkia biglobosa) food Condiment

In order to select starter cultures for controlled fermentation of African locust beans, Bacillus subtilis and B. pumilus from Soumbala were investigated for their ability to degrade arabinogalactan, stachyose, raffinose and sucrose—the main polysaccharides and oligosaccharides in African locust bean. The methods used were liquefaction of gels of galactomannan for screening, pH measurement and High Performance Anion Exhange Chromatography – Pulsed Amperometric Detection (HPAEC–PAD) for detailed degradation study of the carbohydrates. For B. subtilis isolates, liquefaction of galactomannan gels was observed and HPAEC–PAD showed that arabinogalactan and stachyose were partially degraded while raffinose was fully degraded after 48 h of fermentation. Melibiose, fructose and traces of galactose were detected as residual sugars. For B. pumilus isolates much weaker degradation of the carbohydrates was observed. Two isolates of B. subtilis showed strong ability to degrade the oligosaccharides and they are seen as promising starter cultures for fermentation of African locust bean.     

α-Amylases in raw and fermented African oil bean seeds (Pentaclethra macrophylla benth)

The !-amylases of raw and fermented seeds of African oil bean (Pentaclethra macrophylla Benth) were isolated, partially purified (tenfold) and subjected to different thermal and pH conditions. The specific activities of the purified enzyme from raw and fermented seeds were 0.037 ml^-1 min^-1 and 0.88 ml^-1 min^-1 respectively. The !-amylase from fermented seeds was more thermostable with optimum activity at 70 °C, compared to the optimum temperature of 60 °C obtained for the raw seed enzyme. The assayed !-amylases were stable over a wide range of pH (3.0-7.0), with optimum activity found at pH 6.0 and pH 5.0 for raw and fermented seeds respectively. The results show that the !-amylase of Pentaclethra macrophylla complements the microbial amylases in the mainly bacterial fermentation of African oil bean seeds to 'ugba', a highly nutritious food condiment. While the !-amylase in the raw seeds was basically the plant enzyme, that isolated from the fermented seeds was a combination of plant and microbial !-amylases.     

Effects of microencapsulated Allyl isothiocyanate (AITC) on the extension of the shelf-life of Kimchi

Allyl isothiocyanate (AITC) is a well-recognized antimicrobial agent but, application of AITC to food systems is limited due to its high volatility and strong odor. This study was performed to overcome the volatility of AITC by encapsulation using gum Arabic and chitosan and to investigate the effect of microencapsulated AITC as a natural additive on the shelf-life and quality of Kimchi. AITC loaded microparticles were prepared using gum Arabic and chitosan and were added to Kimchi at various concentrations (0-0.02%, w/w). The titratable acidity, pH, microbial changes, and sensory test of Kimchi were examined for 15days at different fermentation temperatures (4 and 10°C). The pH of Kimchi containing AITC microparticles was significantly higher than that of control and the higher the quantity of added AITC, the higher the pH became. The titratable acidity of Kimchi increased during storage especially, titratable acidity of control increased significantly higher than those of Kimchi with added AITC microparticles. The number of Leuconostoc and Lactobacillus species in Kimchi decreased with an increase in the concentration of AITC. The addition of AITC induced reduction of sour taste and improvement of the texture of Kimchi during fermentation. However, as the content of AITC increased, the scores of overall acceptability decreased due to the odor of AITC. These results indicate that addition of AITC (less than 0.1%) to Kimchi is an effective way of enhancing the shelf-life of Kimchi without reducing quality.     

Fermentation Changes the Nutritive Value,Polyphenol Distribution,and Antioxidant Properties of Parkia biglobosa Seeds (African Locust Beans)

This present study was carried out to investigate the effect of natural fermentation on the nutritive value (proximate composition), polyphenolic distribution, and antioxidant properties of African locust beans (Parkia biglobosai). African locust bean (ALB) seeds were boiled for 12 h, dehulled and subjected to natural solid substrate fermentation without any inoculation for 4 days. The proximate composition of the fermented and unfermented locust bean seeds was subsequently determined. Thereafter, the polyphenol distribution, reducing power, DPPH free-radical scavenging antioxidant activity, and inhibition of lipid peroxidation were determined for soluble free and bound polyphenol extracts. The results of the study revealed that fermentation caused a significant increase (P < 0.05) in the ALB protein content with 18.3% in unfermented and 36% in fermented. The fat content in unfermented was 13.2% versus 26.8% in fermented, while there was no significant changes (P > 0.05) in ash, crude fiber, or moisture content. However, there was a significant decrease (P < 0.05) in the carbohydrate content. The total phenolic content, reducing ability and free-radical scavenging ability were high in both the fermented and unfermented ALB. Fermentation caused a significant increase (P < 0.05) in the free phenolic content of the ALB and a significant decrease (P < 0.05) in the bound phenolic content of ALB. The free-phenolic fraction from both fermented and unfermented ALB had high reducing power, free radical scavenging ability, and inhibition of lipid peroxidation using cow brain tissues. Free phenolic extract from the fermented ALB had a significantly higher (P < 0.05) antioxidant activity than that of unfermented ALB. Results suggest that fermentation will increase the nutritive values, free phenolic and antioxidant activity of the African locust bean, making it a potential functional food in addition to its traditional role of dietary protein source.     

Improvement of the traditional processing and fermentation of African oil bean (Pentaclethra macrophylla Bentham) into a food snack--'ugba'

Inocula for the improvement of the traditional production of 'ugba' were developed by attaching pure cultures of Bacillus subtilis responsible for the natural fermentation of the African oil bean seeds on cowpea granules. Changes in pH, amino-nitrogen and protease activity were used as fermentation indicators. In comparison with the natural fermentation, changes in these process variables were more pronounced. Results also showed that the production time could be significantly reduced. In addition, the overall product quality of 'ugba' produced by the developed inocula was good and highly acceptable. The cultures were stable and viable for over 6 months on the granules of cowpea.     

Fermentation of African locust bean (Parkia biglobosa) seeds: involvement of different species of Bacillus

The species of Bacillus which appeared to be responsible for the fermentation of African locust bean seeds were found to be variable. In six separate fermentations, B. subtilis, B. pumilus and B. licheniformis were identified. Bacillus subtilis predominated in four of the fermentations and its growth appeared to be favoured by low oxygen tension. Lactic acid bacteria, Leuconostoc spp., which had been reported in large number and thus implicated in the fermentation by earlier workers were not observed.     

Effect of boiling and roasting on the fermentation of soybeans into dawadawa (soy-dawadawa)

Soybeans which had initially been dehulled by either boiling (boiled/dehulled) or roasting (roasted/dehulled) before peeling, were cooked and fermented into dawadawa, a traditional food condiment. The micropopulation, enzymatic activities, proximate composition, amino acid, and aroma profiles of the two types of soybean dawadawa were evaluated during fermentation. Only minor differences were found in the microbial profiles of the two types of soy-dawadawa. Although boiled/dehulled soy-dawadawa initially had lower microbial counts, it recorded higher counts at the advanced stages of fermentation. Proteolytic and amylolytic Bacillus species including Bacillus subtilis, Bacillus pumilus, Bacillus licheniformis, Bacillus cereus, and Bacillus firmus dominated the micropopulation of the two types of soy-dawadawa with Bacillus subtilis accounting for about 50% of the Bacillus species in all samples. Lactic acid bacteria and yeasts occurred in low numbers in the two types of soy-dawadawa. The proximate composition of the two types of soy-dawadawa were similar, and their contents of moisture and protein increased whilst fat and ash decreased during fermentation. Both types of fermenting soy-dawadawa recorded similar levels of alpha-amylase activity, but boiled/dehulled soy-dawadawa showed slightly higher protease activity. The levels of isoleucine, leucine, lysine, phenylalanine, arginine and proline increased significantly with fermentation time in both types of soy-dawadawa. With respect to differences in their aroma profiles, hexanodecanol, octadecyl acetate, 1,2-dimethyl benzene, tetradecene, (E)-5-eicosene, cyclohexadecane, and hexacosane were found only in the roasted/dehulled samples, whilst 1,2-ethanediol, ethyl acetate, dimethyl disulfide, cyclotetradecane, decene, indole , 2 butyl-octenal, acetophenone, and toluene were found only in the boiled/dehulled samples. A market focus group showed preference for roasted/dehulled soy-dawadawa over boiled/dehulled soy-dawadawa. Apart from the volatile aroma compounds, the biochemical and microbiological profiles of the two types of soy-dawadawa showed only minor differences and were also similar to the profiles reported for African locust bean dawadawa.     

Functional Properties of Flax Seed Mucilage

Flax seed (Linwn usitatissimum L.) mucilage was prepared by extraction of seeds with water followed by evaporation, precipitation with ethanol and freeze drying of extract. Proximate composition, solubility, foamability and moisture sorption characteristics were determined. The mucilage contained less carbohydrates, more minerals and more protein than commercial locust bean and guar gums. Its solubility, however, was higher than locust bean and guar gums, and lower than gum arabic. Flax seed mucilage exhibited good foam stability properties in aqueous solutions at 1.0% (w/v). Very diluted solutions exhibited Newtonian-like behavior while shear thinning was shown at concentrations above 0.2% (w/v). The viscosity was maximum at a pH range 6.0–8.0 and it was reduced in solutions containing NaCl.     

Survival of Listeria monocytogenes and Escherichia coli O157:H7 during sauerkraut fermentation

Sauerkraut was produced from shredded cabbage, as is typical in the United States, and from whole head cabbages, which is a traditional process in parts of Eastern Europe. The sauerkraut was inoculated with five strain mixtures of Escherichia coli O157:H7 and Listeria monocytogenes, and the populations of these bacteria, as well as lactic acid bacteria, pH, and titratable acidity, were monitored over the course of fermentation. Fermentation variables were temperature (18 and 22 degrees C) and salt concentration (1.8, 2.25, and 3%). For most of the analyses, the type of cabbage processing was a significant factor, although within cabbage type, neither salt nor fermentation temperature had significant effects. The final pH of the whole-head sauerkraut was lower than the shredded sauerkraut, but the titratable acidity was significantly higher in the shredded sauerkraut. E. coli O157:H7 and L. monocytogenes persisted in the brines for most of the fermentation, although at the end of the fermentations (15 days for shredded, 28 days for whole head), neither pathogen had detectable populations. E. coli populations decreased more rapidly in the shredded sauerkraut even though the pH was higher because of the higher total acidity in the shredded sauerkraut. Acid-tolerant strains of E. coli and L. monocytogenes were isolated from both shredded and whole-head sauerkraut at different salt concentrations and temperatures after 15 days of fermentation and could be detected at 35 days in the wholehead sauerkraut.     

Determination of shelf life of concentrated yogurt (labneh) produced by in-bag straining of set yogurt using hazard analysis

Strained yogurt, labneh, produced by straining cow's milk set yogurt in cloth bags, was stored at 5, 15, and 25 degrees C, and changes in microbial counts, pH, titratable acidity, percentage of free whey, and sensory attributes were monitored during storage. Counts of total aerobes, psychrotrophic yeasts, yeasts and molds, and lactic acid bacteria, except in samples stored at 25 degrees C, increased irrespective of storage temperature. The pH of samples decreased, titratable acidity and percentage of free whey increased, and texture defects were detected at a later stage than flavor changes during storage. Shelf-life data of labneh was adequately described by the Weibull distribution. The nominal shelf life determined using sensory changes and yeast counts as failure criteria ranged from 8.5 to 10.5, 4.7 to 5.8 and 2.3 to 2.7 d at 5, 15 and 25 degrees C, respectively. Q10 (shelf life at T degrees C/shelf life at T+10 degrees C) for flavor quality loss was 1.98 at 5 degrees C, and the corresponding activation energy was 11.3 kcal/mol.     

Thermally stable hydrogels from enzymatically oxidized polysaccharides

Polysaccharides guar galactomannan (guar gum), locust bean galactomannan (locust bean gum) and tamarind galactoxyloglucan were selectively oxidized by galactose oxidase. The degrees of oxidation of the products were 18-28% for guar galactomannan, 10-16% for locust bean galactomannan and 12-14% for tamarind galactoxyloglucan, calculated from the ratio of oxidized galactose units and total carbohydrates. The rheological properties of the unoxidized and oxidized polysaccharide solutions were investigated by determining their viscosities, storage and loss moduli, and temperature dependence of moduli from 20 °C to 90 °C. All the studied oxidized polysaccharides formed hydrogels throughout the entire temperature range. Concentration (0.2-1% w/v) and degree of oxidation had an effect on the gel formation. The oxidized galactomannans formed stable gels already in low concentrations, such as 0.2-0.4% w/v, while oxidized galactoxyloglucan required a concentration of 0.8% w/v to be stable up to 90 °C. The oxidized polysaccharide hydrogels are highly potential materials for food and medical applications requiring thermal stability.     

Physicochemical, microbiological, and organoleptic profiles of Greek table olives from retail outlets

The physicochemical, microbiological, and organoleptic profile of different commercial table olive products from retail outlets was studied. Average pH values were 4.00, 3.96, and 4.31 for Spanish-style green, naturally black, and dry-salted olives, respectively, while salt content was 6.21, 7.34, and 8.00% for the same commercial products. Mean values for titratable acidity were 0.53 and 0.63% (wt/vol) for green and naturally black olives. In general, mean values for pH, titratable acidity, and salt content were in accordance with the requirements established by the International Olive Oil Council (IOOC) for the trade of table olives, although considerable variation was observed within individual olive samples. Salt content of dry-salted olives did not meet the minimum limit of 10% established by the IOOC. The dominant microbiota consisted of lactic acid bacteria and yeasts. Their population was less than 10(9) CFU ml(-1), as stipulated by the IOOC standard for fermented olives held in bulk in a covering liquid. These microorganisms come from the natural microbiota found in spontaneous fermentations and impose no risk to human health. No enterobacteria, pseudomonads, Bacillus cereus, or Clostridium perfringens were detected in any of the samples given the physicochemical characteristics found. The organoleptic profile varied greatly according to processing style and commercial preparation. Green olives had more uniform sensory characteristics than naturally black and dry-salted olives. The most important attributes that influenced the judgment of the panelists were salt content and crispness of the olives.     

In vitro digestibility of Bacillus fermented soya bean

Bacillus fermented legume products include among others dawadawa and soumbala made from African locust bean, and natto and kinema made from soya bean. Bacillus subtilis is the dominant species involved in the fermentation. During Bacillus fermentation for 48 h of autoclaved soya bean the quantity of soluble and dialyzable matter increased from 22% and 6% up to 65% and 40%, respectively. Protein and carbohydrate degradation during fermentation of soya bean with several Bacillus spp. was investigated and appeared to be substantial during the first 18 h of fermentation resulting in the release of high levels of peptides and oligosaccharides. In vitro digestibility was increased from 29% up to 33–43% after Bacillus fermentation for 48 h. It was shown that Bacillus spp. were able to degrade soya bean macromolecules to a large extent resulting in water-soluble low molecular weight compounds. In vitro digestion of Bacillus fermented soya bean using gastrointestinal enzymes only slightly increased the amount of dialyzable matter, which clearly demonstrated the beneficial effect of Bacillus fermentation on food nutrient availability.     

RHEOLOGICAL STUDY OF O/W EMULSIONS CONTAINING DRIED WHOLE EGG AND LOCUST BEAN GUM

The influence of sucrose addition and type of salt (NaCl; Morton Lite salt: 50% NaCl and 50% KCl, and Mineral salt: 65% NaCl, 25% KCl and 10% MgSO₄) on the rheological properties of O/W emulsions containing locust bean gum was analyzed. Flow curves over the range 0.1 to 100 s⁻¹ and transient flow curves at 0.2 s⁻¹, at 20 and 30C were obtained for 12 different emulsions, containing 30% (w/w) of sunflower oil. The K and n power law parameters and the hysteresis loop area (HL) were evaluated from the flow curve. So were the Hahn parameters from time-dependent stress decay fitting. Rheological behaviour of emulsions was affected by the type of salt and the presence of sucrose. Changes in solvent properties of the continuous phase in the emulsions on the macromolecules present could be responsible for the different behaviour. The ageing effect on flow behaviour parameters and on the hysteresis loop area was also studied. Evolution of rheological parameters during emulsion storage suggests that increase of ionic strength and polar solutes concentration decrease the stabilizing properties of locust bean gum.     

模糊数学感官评价结合响应面法优化低盐荞麦豆酱生产工艺

以苦荞麦和黄豆为原料,制作一款低盐荞麦豆酱。在单因素试验的基础上,以荞麦豆酱的模糊感官评分为响应值,采用Box- Behnken中心组合设计试验对低盐荞麦豆酱的发酵工艺进行优化,并对其产品质量进行分析。结果表明,低盐荞麦豆酱的最优发酵工艺为：发酵时间59 d、荞麦粉添加量30%、发酵温度38 ℃、食盐添加量8%。在此优化条件下,低盐荞麦豆酱的感官评分为（88.49±0.25）分,氨基酸态氮、水分含量分别为（0.78±0.02） g/100 g、（43.22±1.92） g/100 g,其理化和微生物指标均符合相关国标要求。该结果可为低盐荞麦豆酱的开发和生产提供了一定的参考。