Influence of the addition of lupin sourdough with different lactobacilli on dough properties and bread quality

The aim of this research was to study the effects of solid‐state fermentation (SSF) with Lactobacillus sakei, Pediococcus pentosaceus and P. acidilactici on lupine sourdough parameters and lupine sourdough influence on the physical dough properties and wheat bread quality. The results showed that lactic acid bacteria (LAB) significantly reduced the pH and increased total titratable acidity (TTA) of SSF lupine. The highest protease activity in lupine is excreted by L. sakei (187.1 ± 8.6 PU g^?1), and the highest amylase activity, by P. pentosaceus (155.8 ± 7.5 AU g^?1). Lupine sourdough has a significant effect on the rheological properties of doughs, which affect the baking characteristics of the final product. In conclusion, it can be said that L. sakei, P. pentosaceus and P. acidilactici could be used for lupine SSF, and the addition of up to the 10% SSF lupine products increases the wheat–lupine bread quality.     

Comparison of the effect of exopolysaccharide-producing lactic acid bacteria from sourdough on dough characteristics and steamed bread quality

The use of exopolysaccharide (EPS)-producing lactic acid bacteria (LAB) is promising in sourdough fermentation. However, the knowledge of the effects of various species of LAB on steamed bread making remains limited. In this study, the effects of two LAB with high EPS-producing capacity, namely Weissella cibaria L32 and Lactobacillus brevis L17 on dough fermentation and steamed bread quality were estimated. The addition of these two LAB strains significantly increased the titratable acidity and protease activity during the dough fermentation, especially L. brevis L17. Although the in situ EPS synthesised by LAB could improve the steamed bread quality, excessive acidification of L. brevis L17 would still increase the protease activity and thus destroy its gluten network structure. As a result, the steamed bread fermented with L. brevis L17 had the lowest specific volume and hardest texture in comparison with the steamed bread fermented with W. cibaria L32 and with added EPS produced by W. cibaria L32 and L. brevis L17. These results indicated that different EPS-producing LAB exhibited distinctive dough fermentation characteristics, and the in situ EPS-producing W. cibaria L32 could improve steamed bread quality, which confirmed its potential application in steamed bread making.     

Isolation of halophilic lactic acid bacteria from traditional Chinese fermented soybean paste and assessment of the isolates for industrial potential

Lactic acid bacteria (LAB) from traditional Chinese fermented soybean paste were isolated and identified. A total of 61 LAB were selectively obtained from 32 homemade Chinese soybean pastes. The isolated LAB were divided into two groups by their salt tolerance, 28 halophilic LAB and 33 non-halophilic LAB. Phenotypic analysis showed that these LAB belonged to four genera and 13 species. Tetragenococcus halophilus was the predominant species in the identified strains. Four species of LAB were firstly isolated from fermented soybean food product, Lactobacillus panis, Lactobacillus pentosus, Lactobacillus vaccinostercus and Lactobacillus collinoides. T. halophilus T5 showed vigorous growth and fast acidification in high salt concentration. The volatile compounds of mixed microorganism soybean paste with T. halophilus T5, Zygosaccharomyces and Torulopsis candida, during the different fermentation stage were higher in number than those of normal soybean paste at same processing procedure.     

An alkaline protease from Kocuria kristinae F7: properties and characterization of its hydrolysates from soy protein

An alkaline serine protease (SF1) from Kocuria kristinae F7 was purified. The molecular mass of the SF1 protease was estimated to be 57 kDa by sodium dodecyl sulfate–polyacrylamide gel electrophoresis (SDS–PAGE). The N-terminal amino acid sequence of the first 14 amino acids of the SF1 protease showed low homology with bacterial proteases, suggesting that the enzyme had not been described previously. The SF1 protease exhibited maximal activity at pH 9.0 and 60 °C. The activity of the SF1 protease was enhanced by the presence of Ca²⁺ and Mg²⁺ ions. It showed high stability in the presence of NaCl and ethanol. Reverse-phase high-performance liquid chromatography (RP-HPLC) analyses indicated that soybean protein isolates treated with the SF1 protease generated four principal new hydrophilic peptides. Mass spectrometry analyses indicate that the distribution of molecular weight of these peptides was from 0.705 to 1.305 kDa. Hydrolysis of soybean protein isolates with the SF1 protease increased the level of total free amino acids, essential amino acids and flavor amino acids. The SF1 protease may decrease the bitterness of soy protein hydrolysates. The results showed that the SF1 protease of Kocuria kristinae F7 appears to be good candidate enzyme for potential application in acceleration of fermented soybean food ripening.     

Probiotic potential of lactic acid bacteria from fermented Malaysian food or milk products

This study investigated the probiotic characteristics of lactic acid bacteria (LAB) isolated from 14 Malaysian fermented food or milk products. In total, 22.3% (121 of 542) of the LAB isolated from the local fermented products exhibited antibacterial activity against Micrococcus luteus. Twelve LAB that demonstrated better antibacterial activity against M. luteus as compared to a commercial strain Lactobacillus casei strain Shirota (LABPC) were selected for further characterisation. Based on 16S rRNA gene sequence, the LAB were identified as pediococci (seven) and lactobacilli (five). All 12 LAB showed bile tolerance, but only eight were acid tolerant at pH ≥ 3.0. The highest level of adhesion to HT‐29 cells was observed among the Lactobacillus sp. LAB 1 and 10. The LAB also showed antimicrobial activity against Escherichia coli and Staphylococcus aureus through the production of organic acids. LAB isolated from Malaysian fermented food and milk products, especially fermented tapioca, contains potential probiotic candidates.     

Isolation of probiotic bacteria from turmeric (Curcuma longa Linn.) and its application in enriched beverages

The purpose of this study is to obtain lactic acid bacteria (LAB) from fresh turmeric rhizomes that could be used as starter culture with the ability to ferment turmeric and to investigate its influence on total phenolic content (TPC) and antioxidant activity. Three LAB strains, namely Enterococcus faecium, Lactococcus lactis subsp. lactis, and Lactobacillus plantarum, were isolated from turmeric rhizomes. These microorganisms showed similar characteristics as probiotic organisms. The TPC of fermented turmeric beverages was measured using the Folin–Denis reagent method, while the antioxidant activity assays were quantified by 2,2‐diphenyl‐1‐picrylhydrazyl (DPPH) and ferric reducing antioxidant power assay. This study confirms that bacteria isolated from turmeric can be used as starter. Moreover, fermentation time and LAB lead to dominant amount of TPC and that they have high levels of antioxidant activities.     

Investigation of Microbial Communities of Chinese Sourdoughs Using Culture‐Dependent and DGGE Approaches

Sourdough is typically characterized by the complex microbial communities mainly comprising of yeasts and lactic acid bacteria (LAB). The objective of this study was to explore the microbiota of Chinese traditional sourdoughs collected from different areas of China using culture‐dependent and denaturing gradient gel electrophoresis (DGGE) methods. A total of 131 yeasts, 2 molds, and 106 LAB strains were isolated and identified. Based on the culture‐dependent analysis, the populations of yeasts and LAB were at the level of 10⁵ to 10⁷ and 10⁶ to 10⁷ cfu/g, respectively. Similarly, the results of RT‐qPCR showed that the values of total yeasts and LAB populations were in the range of 10⁶ to 10⁷ and 10⁷ to 10⁸ copies/g, respectively. Using culture‐dependent method, a total of 7 yeasts, 2 molds and 7 LAB species were identified. Results showed that Saccharomyces cerevisiae and Lactobacillus plantarum were the predominant species among the yeasts and LAB microflora. Similarly, using PCR‐DGGE approach, 7 yeasts, 1 mold and 9 LAB species were detected. The yeast, S. cerevisiae, represented the predominant, while the yeast Candida tropicalis represented the subdominant species of the yeast community. Among the LAB community, Lactobacillus sanfranciscensis was the predominant species, while Lactococcus qarvieae, Enterococcus faecium, Lactobacillus delbrueckii and Enterococcus cecorum were among the less dominant species.     

Purification and properties of cysteine protease from rhizomes of Curcuma longa (Linn.)

BACKGROUND: Turmeric rhizome (Curcuma domestica Linn.) contains proteases and has proteolytic activity. Curcumin from turmeric rhizomes has been used for healing manu ailments, including cancer have been used for healing many ailments, including cancer. The purpose of this study was to purify turmeric protease and to research their biochemical characteristics. RESULTS: Cysteine protease from C. domestica has been purified to homogeneity using acetone precipitation followed by preparatory native polyacrylamide gel electrophoresis (PAGE). This protocol resulted in six fold purification with 28% final recovery. The purified turmeric protease showed a prominent single peak and band on high‐performance liquid chromatography and sodium dodecyl sulfate–PAGE, respectively, and an estimated molecular weight of 43 KDa, and exhibited optimal activity between 37 and 60 °C. The protease activity of the turmeric protease was significantly inhibited by iodoacetic acid. The turmeric protease had higher alanine and glutamate content and cleaved synthetic peptides N‐Cbz‐Ile‐Pro and N‐Cbz‐Phe‐Leu in a time‐dependent manner. Peptide mass fingerprint using matrix‐assisted laser desorption/ionization–time of flight mass spectroscopy revealed peptide matches to proteasome subunit alpha type 3 of Oryza sativa ssp. japonica (Rice). The turmeric protease showed antifungal activity at 10 µg mL^?1 towards pathogens Pythium aphanidermatum, Trichoderma viride and Fusarium sp. CONCLUSION: Cysteine addition significantly activated turmeric protease. The protease inhibition test suggested that turmeric protease belonged to the cysteine type. The biochemical characteristics of turmeric protease described in this paper can provide useful information for potential end uses of turmeric protease for pharmaceutical industry applications such as therapeutics. Copyright © 2009 Society of Chemical Industry     

Inhibition of Citrus Fungal Pathogens by Using Lactic Acid Bacteria

Abstract: The effect of lactic acid bacteria (LAB) on pathogenic fungi was evaluated and the metabolites involved in the antifungal effect were characterized. Penicillium digitatum (INTA 1 to INTA 7) and Geotrichum citri-aurantii (INTA 8) isolated from decayed lemon from commercial packinghouses were treated with imazalil and guazatine to obtain strains resistant to these fungicides. The most resistant strains (4 fungal strains) were selected for evaluating the antifungal activity of 33 LAB strains, among which only 8 strains gave positive results. The antifungal activity of these LAB strains was related to the production of lactic acid, acetic acid, and phenyllactic acid (PLA). A central composite design and the response surface methodology were used to evaluate the inhibitory effect of the organic acids produced by the LAB cultures. The antifungal activity of lactic acid was directly related to its concentration; however, acetic acid and PLA showed a peak of activity at 52.5 and 0.8 mM, respectively, with inhibition rates similar to those obtained with Serenade^® (3.0 ppm) imazalil (50 ppm) and guazatine (50 ppm). Beyond the peak of activity, a reduction in effectiveness of both acetic acid and PLA was observed. Comparing the inhibition rate of the organic acids, PLA was about 66- and 600-fold more effective than acetic acid and lactic acid, respectively. This study presents evidences on the antifungal effect of selected LAB strains and their end products. Studies are currently being undertaken to evaluate the effectiveness in preventing postharvest diseases on citrus fruits.     

Microbial safety and probiotic potential of packaged yogurt products in Pakistan

Yogurt is a health food with notable market production and demand. Because of this, we conducted a study on prominent commercial brands of yogurts in Pakistan for microbial content and the probiotic potential of the contained lactic acid bacteria (LAB), in the context of their label claims. All contained viable LAB, but the numbers (cfu g⁻¹) varied considerably. Three of the products made explicit probiotic claims, but LAB from these displayed no probiotic attributes per WHO-FAO guidelines. The yogurt starter and nonstarter Lactobacillus strains had no gelatinase or hemolytic activity and exhibited significant antibacterial activity against some human pathogens. One brand with a probiotic claim contained an L. acidophilus strain that showed cholesterol assimilation activity in vitro. Some potential human pathogens that were hemolytic and resistance to β-lactam antibiotics were also detected in the products. The findings demonstrate a need for better quality control and regulation to ensure safety and efficacy of yogurt products.     

Proteolysis characteristics of Actinomucor elegans and Rhizopus oligosporus extracellular proteases under acidic conditions

Enzymatic hydrolysis of soybean protein isolate by the extracellular proteases from Actinomucor elegans and Rhizopus oligosporus at pH 3.0, 3.5, 5.0, 5.5 and 6.0 was investigated. The activity of the A. elegans protease is lower than that of R. oligosporus, but both proteases exhibit considerable degradation of soybean protein at pH 5.5 and 6.0. The water‐soluble protein content and the degree of hydrolysis of the hydrolysates are increased significantly, and bitterness values are very low. Sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS‐PAGE) reveals that these proteases have different cutting sites on peptide polymers. At pH 5.5, there is a lower content of total free amino acids (39.20 mg per 100 mL; 62.68% hydrophobic amino acids) in the R. oligosporus protease hydrolysate. In conclusion, treatment with R. oligosporus protease at pH 5.5 achieves efficient degradation of soybean protein, suggesting a promising industrial process for making bitterless protein hydrolysates.     

Production of biogenic amines by lactic acid bacteria and bifidobacteria isolated from dairy products and beer

The aim was to monitor production of eight biogenic amines (BAs) (histamine, tyramine (TYR), tryptamine, putrescine, cadaverine (CAD), phenylethylamine, spermine and spermidine) by selected 81 lactic acid bacteria (LAB) strains: Lactobacillus, Lactococcus, Leuconostoc, Enterococcus, Pediococcus, Tetragenococcus and Bifidobacterium. The tested LAB and bifidobacteria were isolated from dairy products and beer. The decarboxylase activity of the micro‐organisms was studied in growth medium after cultivation. The activity was monitored by HPLC after the pre‐column derivatisation with dansylchloride. Fifty LAB showed decarboxylase activity. Thirty‐one strains produced low concentrations of CAD (≤10 mg L^?1). Almost 70% of beer isolates generated higher amounts of TYR (≤3000 mg L^?1). Most of the tested LAB demonstrated decarboxylase activity. The above micro‐organisms can contribute to the increase of content of BAs in dairy products or beer and thereby threaten food safety and health of consumers. Production of BAs even by the representatives of some probiotic strains (Bifidobacterium and Lactobacillus rhamnosus) was detected in this research. This study has also proved that contaminating LAB can act as sources of higher amounts of CAD and TYR in beer.     

Improvement of soybean meal quality by one-step fermentation with mixed-culture based on protease activity

Microbial fermentation can improve the quality of soybean meal by reducing the level of anti-nutrition factors and increasing the content of protein and small peptides. In this study, Bacillus velezensis, Enterococcus faecium and Saccharomyces boulardii were used for one-step mixed fermentation of soybean meal. The fermentation of soybean meal was carried out in an anaerobic bag using fermentation liquid of strains with maximum protease activity as inoculum and water. The results of process optimization showed that the mixture ratio of B. velezensis: E. faecium: S. boulardii was 37.5%:25%:37.5%. The three microorganisms added together as a mixture. Under the conditions of 55% inoculation with microorganisms mixture, 0.8% acid protease addition, 46 ℃ and 6 days, the quality of fermented soybean meal was significantly improved. The small peptides increased from 4.48% to 47.4%, the TCA solution protein increased from 44.3 g/kg to 255.2 g/kg, the antigenic protein glycinin decreased by 94.37% from 320.12 μg/mL to 18.02 μg/mL, and β-conglycinin decreased by 85.83% from 232.11 μg/mL to 32.89 μg/mL. Other anti-nutritional factors urease, phytic acid and stachystryose all decreased. The result indicated that it was feasible to improve the feed quality of soybean meal by one-step mixed fermentation based on protease activity.Industrial relevanceFermentation is the main method to improve the feed quality of soybean meal. Our study revealed that one-step mixed-culture fermentation based on microbial protease activity showed a significant effect on increasing beneficial small peptides, reducing anti-nutrient factors and enhancing palatability of soybean meal. This process simplified the operation of mixed fermentation and could be implemented in solid fermentation system of feed and food industry to improve the beneficial properties of products.     

Effect of Lactic Acid Bacteria Fermentation on Rosmarinic Acid and Antioxidant Properties of in vitro Shoot Culture of Orthosiphon aristatus as a Model Study

In vitro shoot culture of Orthosiphon aristatus was selected as a model system for lactic acid bacteria (LAB) fermentation study. This in vitro plant was subjected to different types of LAB fermentations (Lactobacillus plantarum ATCC 8014 and Lactobacillus acidophilus NCFM) for a different time periods of 24, 48, and 72 h at 37^oC. The LAB fermentations consisted of solid state fermentations in a climatic incubator and liquid state of fermentations in a DCU fermenter system. The aim was to determine the effect of fermentation on antioxidant properties of the in vitro plant extract. Results indicate that all types of LAB fermentation decreased the level of rosmarinic acid and total phenolic compounds; however, a slight increase in total flavonoids and flavonols was observed in solid state fermentations samples. The highest reduction was obtained in the sample of liquid state fermentation inoculated with L. plantarum for a period of 72 h. The loss in rosmarinic acid and phenolics was concomitant with a loss of total antioxidant activity (DPPH, TEAC, and SOD-like activity). HPLC result confirmed that the longer fermentation was the greater reduction phenolic acids content was found. These results indicate LAB fermentation caused a decrease on antioxidant properties of in vitro shoot culture of Orthosiphon aristatus.     

Anti-Helicobacter pylori activity of fermented milk with lactic acid bacteria

BACKGROUND: Ten strains of lactic acid bacteria (LAB) were investigated for their anti‐Helicobacter pylori effects. The bactericidal activity and organic acid content in spent culture supernatants (SCS) from fermented milk were measured. In addition, the exclusion effect of SCS against H. pylori infection of human gastric epithelial AGS cells was assayed. RESULTS: Three LAB strains, LY1, LY5 and IF22, showed better anti‐Helicobacter effects than the other strains. There were no significant differences in the bactericidal activity of LAB strains between original SCS, artificial SCS and SCS treated by heating or protease digestion. However, neutralised SCS lost this activity. These results suggest that the anti‐H. pylori activity of SCS may be related to the concentration of organic acids and the pH value but not to protein components. In the AGS cell culture test, both fermented LY5‐SCS and artificial LY5‐SCS significantly reduced H. pylori infection and urease activity (P < 0.05). CONCLUSION: In this study, in vitro methods were used to screen potential probiotics with anti‐H. pylori activity. This may provide an excellent and rapid system for studying probiotics in the functional food and dairy industries. Copyright © 2011 Society of Chemical Industry     

Partial purification and characterisation of cysteine protease in wheat germ

BACKGROUND: Proteases have become an essential part of the modern food and feed industry, being incorporated in a large and diversified range of products for human and animal consumption. The objective of this study was to purify and characterise a protease from wheat germ. RESULTS: After purification a single protease of molecular weight 61–63 kDa (determined by sodium dodecyl sulfate polyacrylamide gel electrophoresis) was obtained. The purified protease had optimal activity at 50 °C and maintained its activity completely after incubation at 30 °C for 30 min, while over 47% of the activity was lost after incubation at 80 °C for 30 min. The purified protease had optimal activity and maintained maximum stability at pH 5.5, while the activity decreased after incubation for 30 min at other pH values. The protease was inhibited by Mg²⁺, Mn²⁺, Ba²⁺ and iodacetic acid and stimulated by Li⁺, Ca²⁺, Cu²⁺, β‐mercaptoethanol and dithiothreitol, while Zn²⁺, L ‐cysteine and glutathione had no significant effect on its activity. At pH 5.5 the enzyme had a K_m of 0.562 mg mL^?1 with casein as substrate and showed higher affinity to casein than to bovine serum albumin, ovalbumin and gelatin. CONCLUSION: The purified enzyme from wheat germ was identified as a cysteine protease. Copyright © 2011 Society of Chemical Industry     

Relationships between the bacterial diversity and metabolites of a Chinese fermented pork product,sour meat

The present study evaluated the relationship between the bacterial diversity and metabolites of various sour meats, a traditional fermented meat product. The results showed that sour meats had a low pH (average of 4.34), low water activity (a_w, an average of 0.917), and high lactic acid (~19 177.48 mg kg⁻¹) and free amino acid (FAA) (~4047.65 mg kg⁻¹) concentrations. The lactic acid bacteria (LAB), especially Lactobacillus and Weissella, were predominated in the samples. The alcohols, aldehydes and esters accounted for the major components of volatile organic compounds (VOCs). Of which, (E)-2-octenal, nerol, cis-4-decenal, tetradecanal, linalyl acetate, linalool, 1-octen-3-ol, hexanal, 1-heptanol, benzaldehyde and (E)-2-decenal were considered as the key VOCs of sour meats. The redundancy analysis (RDA) and Pearson’s correlation analysis revealed that LAB (Lactobacillus and Weissella) and salt were positively correlated with most FAAs and VOCs. Overall, higher salinity, lower a_w and abundant LAB might be the indicators of good sour meat quality.     

Purification and characterization of Aspergillus oryzae LK-101 salt-tolerant acid protease isolated from soybean paste

A novel salt-tolerant acid protease was produced from Aspergillus oryzae LK-101 (AOLK-101). The AOLK-101 protease was purified to homogeneity by ammonium sulfate precipitation, DEAE-Sephadex A-50 and Sephadex G-100 chromatographies in order. The specific activity and the purification ratio of the purified protease were 2,301 unit/mg and 11.6 fold, respectively, with 25 kDa of molecular weight on sodium dodecyl sulfate-polyacrylamide gel electrpphoresis (SDS-PAGE). Its optimal pH and temperature were pH 6.5 and 50°C, respectively. This protease was relatively stable at pH 4.5–7.5, below 40°C, and up to 10% salt concentration. The protease was moderately inhibited by Ag²⁺ and Zn²⁺, and strongly by ethylenediamide tetraacetic acid (EDTA) and phenylmethysulfonyl fluoride (PMSF), but activated by Cu²⁺ and Mn²⁺. Therefore, the AOLK-101 protease was a serine protease based on the influence of metal ions and inhibitors. K _m , V _max , k _cat , and k _cat /K _m values of AOLK-101 protease for hammastein milk casein were 1.04 mg/mL, 124.84 unit/L, 163.5/sec, and 3.9×10⁶/m·sec, respectively.     

Control of lactic acid bacteria in fermented beverages using lysozyme and nisin: test of traditional beverage boza as a model food system

The objective of this study was to increase quality and limited shelf‐life of boza (3–15 days), a traditional Balkan origin fermented beverage using lysozyme (LYS) and/or nisin (NIS). For this purpose, the effectiveness of NIS, LYS and LYS:NIS combinations was first tested in a broth medium at 4 °C for 3 weeks on Lactobacillus plantarum, one of the frequently isolated lactic acid bacteria in boza. Stability of LYS and NIS in boza, their effects on LAB counts, and chemical and sensory properties of boza were then evaluated during cold storage at 4 °C. Results of LAB counts as well as pH, d ‐ and l ‐lactic acid, and titratable acidity measurements showed that LAB in boza containing NIS (250 μg g^?1) or LYS:NIS (500:250 μg g^?1) could be controlled without reducing LAB counts below 6 log CFU mL^?1 during 2 weeks shelf‐life. In contrast, LYS (500 μg g^?1) alone could not control LAB in boza to delay its acidic spoilage. Positive effects of NIS and LYS:NIS application on quality of boza were also proved with sensory analysis by panelists and e‐nose measurements. This work showed that use of natural GRAS agents in preservation of fermented beverages containing probiotic LAB is possible without affecting their characteristic aroma and flavour.     

The proteolytic activity of selected lactic acid bacteria in fermenting cow's and camel's milk and the resultant sensory characteristics of the products

The aim of this study was to evaluate the proteolytic activities of 14 strains of lactic acid bacteria and their impact on sensory characteristics of the resultant fermented cow and camel milk. The results showed that Lactobacillus rhamnosus PTCC 1637 and Lactobacillus fermentum PTCC 1638 had high protease activity and high mean values for sensory quality in fermented cow's and camel's milk. Lactobacillus plantarum PTCC 1058 revealed high protease activity and sensory scores in camel milk. Consequently, milk fermentation using such strains might be recommended for the development of dairy products containing bioactive peptides.