Free D- and L-Amino Acid Evolution During Sourdough Fermentation and Baking

The evolution of free D- and L-amino acids in sourdoughs started with various lactic acid bacteria (LAB) and yeasts was studied. Lactobacillus brevis subsp. lindneri CB1 and Lactobacillus plantrum DC400 had high proteolytic activity. During sourdough fermentation, Saccharomyces cerevisiae 141 and Saccharomyces exiguus M14 sequentially utilized free amino acids produced by bacterial activity. Due to increased cell yeast autolysis, more S. exiguus M14 inocula caused more free amino acids which were partially utilized by LAB without causing hydrolysis of wheat flour protein. D-alanine, D-glutamic acid and traces of other D-isomers were observed in sourdoughs fermented with L. brevis subsp. lindneri CB1 and S. cerevisiae 141. Free total D- and L-amino acid content decreased by more than 44% after baking the sourdoughs. No abiotic generation of new D-amino acid isomers was detected in the baked sourdoughs.     

乳酸菌发酵对燕麦淀粉物化及热力学特性的影响

以燕麦粉为原料,分别利用植物乳杆菌(L.p)和旧金山乳杆菌(L.s)两种乳酸菌对其进行发酵,研究燕麦淀粉在发酵过程中各种物化及热力学特性的变化。结果表明：燕麦粉经过两种菌发酵后pH 值下降,且L.p 发酵的燕麦粉pH 值下降的速率和产酸量都大于L.s。但是到发酵后期,两种乳酸菌发酵的燕麦粉pH 值相近。发酵燕麦淀粉的溶解度和溶胀力都随着温度的升高而增加,在不同温度下,其溶胀力和溶解度在发酵过程中的变化趋势不同,经L.P 发酵的样品的溶胀力低于L.s,但溶解度大于经L.s 发酵的样品。快速黏度分析仪(RVA)和差示扫描量热仪(DSC)分析得到发酵过程中燕麦淀粉的变化：发酵后燕麦淀粉糊化过程中峰值黏度随着发酵时间的延长而降低,糊化起始温度提前,糊化所需时间延长,糊化焓升高,且L.s 发酵样品的糊化焓值高于L.p。发酵后燕麦淀粉的直链淀粉含量在发酵过程中呈上升趋势,L.p 发酵的样品的直链淀粉的含量高于L.s。     

恩施鲊广椒乳酸菌的分离鉴定及其对挥发性风味物质的影响

采用传统培养方法对采集自湖北恩施地区的15份鲊广椒样品中的乳酸菌进行分离,采用分子生物学技术对其进行鉴定。然后,采用优势乳酸菌制作鲊广椒,并利用电子鼻技术分析其挥发性风味物质。最后,通过主成分分析（PCA）筛选出适合鲊广椒发酵的乳酸菌菌种。结果表明,从鲊广椒样品中共分离鉴定出植物乳杆菌（Lactobacillus plantarum）10株、发酵乳杆菌（Lactobacillus fermentum）3株、副干酪乳杆菌（Lactobacillus paracasei）1株和食品乳杆菌（Lactobacillus alimentarius）1株,其中,L. plantarum占总分离菌株的66.67%,是湖北恩施鲊广椒中的优势乳酸菌。电子鼻分析结果表明,与对照相比,添加L. plantarum的鲊广椒芳香类物质明显增多,氮氧化物、氢化物、有机硫化合物和烷烃等物质含量显著降低。PCA结果表明,L. plantarum HUBAS51132和HUBAS51141制作的鲊广椒其芳香性更好。     

Use of starter cultures of lactic acid bacteria and yeasts in the preparation of togwa,a Tanzanian fermented food

Starter cultures of lactic acid bacteria (Lactobacillus brevis, Lactobacillus cellobiosus, Lactobacillus fermentum, Lactobacillus plantarum and Pediococcus pentosaceus) and yeasts (Candida pelliculosa, Candida tropicalis, Issatchenkia orientalis and Saccharomyes cerevisiae) isolated from native togwa were tested singly or in combination for their ability to ferment maize-sorghum gruel to produce togwa. All species of bacteria showed an ability to ferment the gruel as judged by lowering the pH from 5.87 to 3.24-3.49 and increasing the titratable acidity from 0.08% to 0.30-0.44% (w/w, lactic acid) in 24 h. Yeasts used singly showed little activity within 12 h, but lowered the pH to 3.57-4.81 and increased the acidity to 0.11-0.21% in 24 h. Yeasts in co-culture with lactic acid bacteria (LAB) had a modest effect on the final acidity (P<0.05). The number of lactic acid bacteria and yeasts increased while the Enterobacteriaceae decreased with fermentation time. The pH was lowered and lactic acid produced significantly (P<0.05) fastest in natural togwa fermentation and in samples fermented by L. plantarum or L. plantarum in co-culture with I. orientalis. The content of fermentable sugars was reduced during fermentation. Most volatile flavour compounds were produced in samples from fermentation by P. pentosaceus and I. orientalis in co-culture with either L. plantarum or L. brevis.     

协同发酵在植物乳杆菌发酵乳中的应用研究进展

植物乳杆菌由于其益生功能受到广泛关注。植物乳杆菌蛋白水解系统不完全,存在肽转运系统和胞内肽酶,但缺乏胞壁蛋白酶,这导致该菌无法直接利用乳中蛋白质,因此在乳中生长不佳。利用蛋白酶活性强的乳酸菌和植物乳杆菌的协同发酵是一种解决方法,前者能初步水解乳中蛋白质产生多肽和游离氨基酸,为植物乳杆菌提供氮源,促进其生长。本文综述了植物乳杆菌益生功能,在乳中生长不良的原因及相关解决办法(添加营养物质、添加蛋白酶、协同发酵)的研究进展。     

四川泡菜中产γ-氨基丁酸微生物的系统发育与表达能力评估

为拓展产γ-氨基丁酸(γ-aminobutyric,GABA)微生物资源,以四川泡菜为分离源,从中分离产GABA的乳酸菌和酵母,并对其GABA表达能力进行评估。通过分离纯化,从四川泡菜中获得了338株乳酸菌和67株酵母。采用纸色谱测定,筛选获得12株乳酸菌和3株酵母菌具有产GABA的能力。生理生化和系统发育研究揭示12株乳酸菌分别被鉴定为Lactobacillus acidophilus(占比8.3%)、Lactobacillus fermentum(8.3%)、Lactobacillus kimchii(8.3%)、Lactobacillus suebicus(8.3%)、Lactobacillus brevis(16.7%)、Lactobacillus parafarraginis(8.3%)、Lactobacillus similis(8.3%)、Lactobacillus plantarum(25.2%)和Lactobacillus pentosus(8.3%);3株酵母中,2株被鉴定为Saccharomyces cerevisiae;1株被鉴定为Candida tanzawaensis。进一步采用高效液相色谱对菌株GABA的表达能力评估发现,乳酸菌Lactobacillus plantarum BC114和Lactobacillus brevis BC237发酵产GABA的能力较强,分别达1 720 mg/L和1 080 mg/L;酵母菌Saccharomyces cerevisiae JM037发酵产GABA的能力较强,达670 mg/L。     

酸面团中优势乳酸菌和酵母菌对麦谷蛋白结构的影响

以分离自我国传统酸面团中的优势菌种植物乳杆菌（Lactobacillus plantarum）Sx9、旧金山乳杆菌（Lactobacillus sanfranciscensis）Gm4及酿酒酵母（Saccharomyces cerevisiae）Sq7为发酵菌种,研究其对小麦麦谷蛋白大聚体的溶解性、游离巯基含量及二级结构变化的影响。结果表明,发酵24 h后,植物乳杆菌Sx9、旧金山乳杆菌Gm4及酿酒酵母Sq7使麦谷蛋白大聚体的溶解率分别增加7.06%、2.47%、2.47%。在发酵6～12 h,植物乳杆菌Sx9对二硫键作用明显,发酵液中游离巯基含量从0.29 μmol/mg迅速增加至0.61 μmol/mg。植物乳杆菌Sx9和旧金山乳杆菌Gm4对二级结构的影响明显,发酵6 h后,无规则卷曲含量几乎为零,全部转化为β-折叠和β-转角,而酿酒酵母Sq7影响甚微,仅有极少量的转化。因此,相比于酿酒酵母Sq7,植物乳杆菌Sx9和旧金山乳杆菌Gm4对麦谷蛋白大聚体的降解效果较好,且植物乳杆菌Sx9最佳。     

Proteolytic activity of lactic acid bacteria strains and fungal biota for potential use as starter cultures in dry-cured ham

During the processing of dry-cured meat products, sarcoplasmic and myofibrillar proteins undergo proteolysis, which has a marked effect on product flavor. Microbial proteolytic activity is due to the action of mostly lactic acid bacteria (LAB) and to a lesser extent micrococci. The proteolytic capacity of molds in various meat products is of interest to meat processors in the Mediterranean area. Eleven LAB and mold strains from different commercial origins were tested for proteolytic activity against pork myosin, with a view to possible use of these strains as starter cultures for Iberian dry-cured ham. Proteolytic activity was tested by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The LAB strains with the highest proteolytic activity were Lactobacillus plantarum (L115), Pediococcus pentosaceus (Saga P TM), and Lactobacillus acidophilus (FARGO 606 TM). The best fungal candidate was Penicillium nalgiovense LEM 50I followed by Penicillium digitatum, Debaryomyces hansenii, and Penicillium chrysogenum.     

Use of selected sourdough strains of Lactobacillus for removing gluten and enhancing the nutritional properties of gluten-free bread

Forty-six strains of sourdough lactic acid bacteria were screened for proteolytic activity and acidification rate in gluten-free (GF) flours. The sourdough cultures consisted of Lactobacillus sanfranciscensis LS40 and LS41 and Lactobacillus plantarum CF1 and were selected and used for the manufacture of GF bread. Fermentation occurred in two steps: (i) long-time fermentation (16 h) and (ii) fast fermentation (1.5 h) using the previous fermented sourdough as inoculum (ca. 43%, wt/wt) with Saccharomyces cerevisiae (baker's yeast). GF bread started with baker's yeast alone was used as the control. Gluten was added to ingredients before fermentation to simulate contamination. Initial gluten concentration of 400 ppm was degraded to below 20 ppm only in the sourdough GF bread. Before baking, sourdough GF bread showed phytase activity ca. sixfold higher than that of GF bread started with baker's yeast alone. Atomic absorption spectrophotometric analysis revealed that the higher phytase activity resulted in an increased availability of free Ca2+, Zn2+, and Mg2+. The concentration of free amino acids also was the highest in sourdough GF bread. Sourdough GF bread had a higher specific volume and was less firm than GF bread started with baker's yeast alone. This study highlighted the use of selected sourdough cultures to eliminate risks of contamination by gluten and to enhance the nutritional properties of GF bread.     

Parameters of rye,wheat, barley,and oat sourdoughs fermented with Lactobacillus plantarum LUHS135 that influence the quality of mixed rye–wheat bread,including acrylamide formation

A Lactobacillus plantarum strain was used for the production of rye, wheat, barley, and oat sourdoughs, and the influence of different sourdoughs on mixed rye–wheat bread quality parameters and acrylamide formation was evaluated. L. plantarum LUHS135 demonstrated versatile carbohydrate metabolism, good growth and acidification rates, and the ability to excrete amylolytic and proteolytic enzymes in various cereal sourdoughs. The same starter and different cereal substrates allow to produce sourdoughs showing different characteristics. The type of sourdough and its quantity had significant influence on acrylamide content in bread (P ≤ 0.0001), and using 5% or 10% of wheat sourdough, 5%, 15%, or 20% of barley sourdough, and 5% or 15% of oat sourdough, it is possible to reduce acrylamide content in bread. Thus, manufacturers need to take into account application of apparent technological approaches for acrylamide in bread reducing.     

四川牦牛奶和曲拉中九株乳酸菌的分子鉴定

为准确鉴定分离自我国四川鲜牦牛奶和曲拉中9株乳酸菌到种水平,作者运用16S rDNA序列分析、recA基因特异扩增和hsp60-RFLP等多种分子技术对分离自我国四川地区鲜牦牛奶和曲拉中的9株乳酸菌进行分类鉴定。结果证实,16S rDNA序列分析法可将9株乳酸菌初步归类为植物乳杆菌群(4株),肠膜明串珠菌(4株),瑞士乳杆菌(1株)。由于16S rDNA序列分析法不能区分植物乳杆菌和戊糖乳杆菌,为了进一步鉴定植物乳杆菌群中的4株菌,继续采用recA基因特异扩增和hsp60-RFLP技术对其细分,结果表明recA基因特异扩增和hsp60-RFLP的方法均能很好地把植物乳杆菌群中的4株菌鉴定到种水平,且均为植物乳杆菌。     

乳酸菌和酵母菌发酵红枣汁工艺优化及成分分析

以红枣汁为原料,探究酿酒酵母分别与发酵乳杆菌、干酪乳杆菌、短乳杆菌和植物乳杆菌复合发酵对红枣汁品质的影响,并确定发酵工艺条件,以期得到一种新型红枣乳酸发酵饮品。通过单因素试验,在37℃条件下发酵60 h,对红枣汁发酵液的总酸度、总酚含量和抗氧化能力进行测定,确定酿酒酵母和乳酸菌的最佳组合及最佳接种比例。结果表明:酿酒酵母与发酵乳杆菌的接种体积比例为1∶4时,红枣汁发酵液总酸度为9.897 g/L,产酸率为8.31%,总酚含量为287.81 mg/L,维生素C含量为6.83 mg/100 mL,2,2′-联氮-双-3-乙基苯并噻唑啉-6-磺酸[2,2′-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid),ABTS]阳离子自由基清除率、1,1-二苯基-2-苦基苯肼(1,1-diphenyl-2-picrylhydrazyl,DPPH)自由基清除率和羟基自由基清除率分别为87.37%、89.38%和76.12%;经酿酒酵母和发酵乳杆菌复合发酵的红枣汁品质优于发酵乳杆菌发酵的红枣汁品质及未发酵的红枣汁;发酵后,发酵液中乳酸、柠檬酸、乙酸和富马酸含量显著增加,草酸、苹果酸、酒石酸和琥珀酸变化不明显;氨基酸中的鲜味氨基酸和甜味氨基酸含量增加明显,表明红枣汁品质得到提升。     

产乳化活性多糖乳酸菌的筛选及其发酵荞麦酸面团、面包的特性

本研究从不同来源的原料中分离得到18株具有产胞外多糖(EPS)能力的乳酸菌,通过对比发酵液及乳酸菌荞麦酸面团的乳化活性,获得一株产EPS且乳化活性较高的乳酸菌,并应用于荞麦酸面团的制作。采用动态流变测定、激光共聚焦观察、质构分析等手段,研究其对荞麦酸面团面包烘焙学特性的影响。结果表明:优选产乳化性多糖的乳酸菌YC'-22经鉴定为发酵乳杆菌(Lactobacillus fermentum strain HBUAS54017),且同源性为100%,EPS产量为1.63 g/L。通过添加乳酸菌荞麦酸面团,使面包面团的面筋网络结构得到强化。相比空白面包(CB),添加1.0%阿拉伯胶的荞麦面包(AGB)、荞麦酸面团面包(SDB)的比容分别增加了12.42%和14.08%,硬度分别降低了13.90%和22.87%;其中SDB面包整体可接受度最高,改善效果最明显。因此,产乳化性多糖的乳酸菌发酵的荞麦酸面团可以作为一种天然的面包功能配料。     

Use of Lactobacillus strains to start cassava fermentations for Gari production

Two Lactobacillus strains, Lactobacillus plantarum BFE 6710 and Lactobacillus fermentum BFE 6620, were used to start cassava fermentations in a pilot study under field production conditions in Kenya, to determine their potential to establish themselves as predominant lactobacilli during the fermentation. Predominant strains from three fermentations were isolated throughout the 48 h fermentation period. The use of these strains in high numbers clearly resulted in 1 to 2 log higher lactic acid bacteria (LAB) counts over the course of the fermentation when compared to the uninoculated control. 178 predominant LAB isolates were grouped based on their phenotypic characteristics, and were characterised to strain level by RAPD-PCR, followed by PFGE strain typing. Overall, L. plantarum strains represented the majority of the isolates, followed by Weissella confusa and Lactococcus garvieae strains. The results of RAPD-PCR and PFGE strain typing techniques indicated that L. plantarum BFE 6710 was successful in asserting itself as a predominant strain. In contrast, L. fermentum BFE 6620 failed to establish itself as a predominant organism in the fermentation. The success of the L. plantarum strains to predominate in the cassava fermentation demonstrates the potential for development of Lactobacillus starter cultures to industrialise the Gari production process.     

Influence of Monoglycerides of Different Chain Lengths on Texture and Flavor of Breads Made with Waxy Cornstarch

The addition of monoglycerides (MGs) monolaurin (ML), monomyristin (MM), monopalmitin (MP), and monostearin (MS)) to a model system bread made from waxy cornstarch was studied for their effects on delaying staling. The model system breads containing 0.5% of a MG and a control bread containing no MG were prepared five different times and stored at room temperature for 1, 3, and 6 days. A sensory panel measured changes in firmness, stale flavor, and moistness, and an Instron universal testing machine (model 1122) was used to indicate firmness of the samples. The volumes of the loaves were measured on day 1 for the five replications. Results indicated that the addition of MGs to the model system breads decreased firmness and stale flavor and increased moistness compared with the control bread. Breads containing MM and MP were rated significantly (p<0.05) less firm and less stale on days 1 and 6 and more moist on day 1 than the breads containing MS and the control bread. In general, as the molecular weight (MW) of the MGs increased, the firmness (by Instron) of the breads also increased, and the volume of the loaves decreased. The control bread had the highest Instron firmness value and the lowest volume when compared with the breads containing MGs.     

Use of sourdough to reduce phytic acid and improve zinc bioavailability of a traditional flat bread (sangak) from Iran

In this study, effects of 8 different sourdough starters prepared with Saccharomyces cerevisiae, Lactobacillus plantarum, L. acidophilus, and Leuconostoc mesenteroides were investigated on the phytic acid level and mole ratio of phytic acid to zinc in a traditional Iranian bread (sangak). Different sourdough preparations were made and incubated at 30°C for 16 h and added to the dough at 10, 20, and 30% replacement levels. Use of sourdough resulted in a decrease in phytic acid level (also in a decrease in the phytic acid to zinc mole ratio) and corresponding increase in zinc bioavailability index when compared to the commercial sangak bread. The lowest phytic acid concentration and highest zinc bioavailability index were achieved when S. cerevisiae, L. plantarum, and Leu. mesenteroides were used at 30.0% dough replacement with sourdough. This study provides awareness about the negative impacts of higher phytic acid level in the breads, which is particularly the case for sangak bread, and also provides a solution for such issue.     

Carbohydrate, peptide and lipid metabolism of lactic acid bacteria in sourdough

The metabolic pathways of lactic acid bacteria that influence bread quality are coupled to the central carbon flux by the availability of cofactors influencing the cellular and environmental redox potential. Homo- and heterofermentative metabolism differ fundamentally with respect to the requirement for regeneration of reduced cofactors, NADH or NADPH. The utilization of co-substrates such as oxygen or fructose as electron acceptors by obligate heterofermentative lactobacilli is coupled to an increased production of acetate in dough. Recently, several oxidoreductases involved in cofactor regeneration were characterized and glutathione and short-chain aldehydes derived from lipid oxidation were identified as substrates for cofactor regeneration by Lactobacillus sanfranciscensis. Based on the different metabolic requirements for cofactor regeneration, homo- and heterofermentative lactobacilli exert divergent effects on redox-reactions in sourdough that influence bread quality beyond the formation of acetate. Proteolysis, followed by peptide or amino acid metabolism by LAB is one of the key routes of flavour formation in bread flavour, and enables the strain-specific formation of antifungal metabolites. Peptide metabolism as well as the metabolism of cysteine, arginine, and phenylalanine in Lactobacillus plantarum, L. sanfranciscensis, and Lactobacillus pontis is increasingly understood and these insights provide new opportunities for the directed application of sourdough LAB for improved bread quality.     

植物乳杆菌发酵紫薯粉对酸面团面包的抗氧化特性及品质影响

采用紫薯粉和植物乳杆菌发酵紫薯粉制作富含花青素的面包,通过对比研究乳酸菌发酵紫薯粉对面包品质的影响;并探讨乳酸菌发酵紫薯粉对面包中花青素含量、总酚含量以及DPPH自由基清除率的影响。结果表明：相比紫薯粉面包,乳酸菌发酵紫薯粉使得面包pH值降低,总酸度(TTA)增加,面包比容有所减小,面包色度C*值增加而色相H*值减小,面包色泽由原来的浅紫色变为浅红色。感官评定的结果显示,采用紫薯粉或乳酸菌发酵紫薯粉制作的面包都为消费者所喜爱。同时,与紫薯粉面包相比,乳酸菌发酵紫薯粉面包的总酚含量和DPPH自由基清除率分别增加了90.8%和6.1%,增强了面包的抗氧化性。     

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.     

On the microbiological profile of traditional Portuguese sourdough

Traditional manufacture of bread from maize has been noted to play important roles from both economic and social standpoints; however, enforcement of increasingly strict hygiene standards requires thorough knowledge of the adventitious microbiota of the departing dough. To this goal, sourdough as well as maize and rye flours from several geographic locations and in two different periods within the agricultural year were assayed for their microbiota in sequential steps of quantification and identification. More than 400 strains were isolated and taxonomic differentiation between them was via Biomerieux API galleries (375 of which were successfully identified) following preliminary biochemical and morphological screening. The dominant groups were yeasts and lactic acid bacteria (LAB). The most frequently isolated yeasts were Saccharomyces cerevisiae and Candida pelliculosa. The most frequently isolated LAB were (heterofermentative) Leuconostoc spp. and (homofermentative) Lactobacillus spp.; L. brevis, L. curvatus, and L. lactis ssp. lactis were the dominant species for the Lactobacillus genera; Lactococcus lactis ssp. lactis for lactococci; Enterococcus casseliflavus, E. durans, and E. faecium for enterococci; and Streptococcus constellantus and S. equinus for streptococci.