Production of lactic acid from vine‐trimming wastes and viticulture lees using a simultaneous saccharification fermentation method

An effective process for the chemical–biotechnological utilization of trimming wastes of vineshoots, an agricultural waste with little use, is reported. Initial treatment with sulfuric acid (prehydrolysis) allowed the solubilization of hemicelluloses to give xylose and glucose‐containing liquors (suitable to make fermentation media for lactic acid production with Lactobacillus pentosus) and a solid phase containing cellulose and lignin. The solid residues from prehydrolysis were treated with NaOH in order to increase their cellulase digestibility. In the alkaline treatments, the effects of temperature (in the range, 50–130 °C), reaction time (30–120 min) and NaOH concentration (4–12 wt% of solution) on the composition and susceptibility to enzymatic hydrolysis of solid residues were assessed by means of an experimental plan with factorial structure. The lignin content decreased, whereas the susceptibility towards the enzymatic hydrolysis increased with temperature, reaction time and NaOH concentration within the tested range. Using the cellulosic residues achieved under the harsher conditions, favorable fermentation kinetics during simultaneous saccharification and fermentation carried out by L rhamnosus for lactic acid production were observed. The nutrients employed were the complete MRS broth and a cheaper medium developed using viticulture lees coming from the white wine making technology. In all cases the final lactic acid concentration achieved was similar, although the volumetric productivity was lower when using lees due to inhibitory effects over the enzymatic hydrolysis. Copyright © 2004 Society of Chemical Industry     

Simultaneous lactic acid and xylitol production from vine trimming wastes

Hemicellulosic hydrolyzates from vineshoot trimmings obtained by dilute sulfuric acid hydrolysis were evaluated for xylitol production by Debaryomyces hansenii NRRL Y‐7426. Bioconversion was not efficient, however, since a mixture of products (mainly ethanol) was achieved. Taking into account that hexoses (such as glucose or mannose) can inhibit xylose metabolism by repression and inactivation of the xylose transport system or catabolic enzymes and that these hemicellulosic hydrolyzates are characterized by a high glucose concentration, a novel technology was developed, sequentially transforming glucose into lactic acid by Lactobacillus rhamnosus followed by fermentation of xylose into xylitol by Debaryomyces hansenii after L. rhamnosus removal by microfiltration. Optimal conditions were achieved using detoxified concentrated hemicellulosic hydrolyzates, after CaCO₃ addition in both stages of fermentation and using nitrogen purges after sampling in order to reduce the oxygen dissolved. Under these conditions 31.5 g lactic acid L^?1 (Q_LA = 1.312 g L^?1 h^?1 and Y_LA/S = 0.93 g g^?1) and 27.5 g xylitol L^?1 (Q_Xylitol = 0.458 g L^?1 h^?1 and Y_Xylitol/S = 0.53 g g^?1) were produced. Finally, lactic acid was selectively recovered using the resin Amberlite IRA 400 (0.0374 g of lactic acid g^?1 of dry resin), allowing a further recovery of xylitol by sequential stages of adsorption, concentration, ethanol precipitation, concentration and crystallization to obtain food‐grade xylitol according to a developed process. Copyright © 2007 Society of Chemical Industry     

Induced lactic acid fermentation of untreated green olives of the Conservolea cultivar by Lactobacillus pentosus

Inoculation of untreated green olives of the Conservolea cultivar (Olea europea media rotunda) with a commercial strain of Lactobacillus pentosus with/without glucose supplement was studied. Despite an initial loss in viability of 0.5 log cycles on average, due to lack of adaptation of the starter to the saline environment of the brine, cultures grew well and initiated an accelerated fermentation process. Inoculation reduced the survival period of Enterobacteriaceae, and consequently potential spoilage, and caused a quicker acidification of brines and decrease in pH compared with control uninoculated processes. The diffusion of phenolic compounds was slow, hindered by the epidermis of the fruits, and had no effect on the growth of the starter. HPLC analysis revealed that lactic and acetic acids were among the end‐products during the fermentation of green olives. Citric, tartaric and malic acids were also detected but at much lower concentrations. Results obtained from this work can be applied directly to industry for the effective use of starter cultures on natural fermentation processes of green olives without prior lye treatment. © 2003 Society of Chemical Industry     

Revalorisation of vine trimming wastes using Lactobacillus acidophilus and Debaryomyces hansenii

BACKGROUND: Mild acid treatments of vine‐shoot trimmings result in the hydrolysis of hemicellulosic sugars that can be utilised by Lactobacillus acidophilus CECT‐4179 (ATCC 832) and Debaryomyces hansenii NRRL Y‐7426 as carbon sources to obtain food additives. Since the high content of glucose in these hydrolysates reduces the effective bioconversion of xylose into xylitol by D. hansenii, the use of Lactobacillus acidophilus, one of the main probiotic species, allows this problem to be solved by the selective consumption of glucose. In order to use both sugars (glucose and xylose), hemicellulosic vine‐shoot trimming hydrolysates can be sequentially fermented by both micro‐organisms. RESULTS: It was found that, in the first step, L. acidophilus generated almost exclusively lactic acid (32.7 g of lactic acid L^?1, Q_LA = 1.363 g L^?1 h^?1, Y_LA/S = 0.72 g g^?1) by homofermentative degradation of sugars (mainly glucose), and in the second step, the remaining hemicellulosic sugars were transformed primarily into xylitol by Debaryomyces hansenii (31.3 g of xylitol L^?1, Q_Xylitol = 0.708 g L^?1 h^?1, Y_Xylitol/S = 0.66 g g^?1). Furthermore, L. acidophilus proved to be a strong cell‐bounded biosurfactant producer. Cell extracts were able to reduce the surface tension (ST) of PBS in 18 mN m^?1 units. Lactobacillus acidophilus cells showed no difference in viability before or after PBS extraction of biosurfactants, achieving values of 0.9 × 10⁹ colony‐forming units (CFU) mL^?1 in both cases. CONCLUSIONS: These results have made a serious contribution to the re‐evaluation of a useless and pollutant residue, producing a wide range of natural food additives. Copyright © 2008 Society of Chemical Industry     

戊糖乳杆菌的鉴定及其发酵D-乳酸的研究

分离得到1株乳酸杆菌LCA,根据其形态、生理生化性质和16S rDNA分子生物技术鉴定,被鉴定为戊糖乳杆菌（Lactobacillus pentosus）,其发酵产物经手性柱高效液相色谱法测定,含有光学纯度为80%以上的D-乳酸,并能利用D-木糖产生D-乳酸。筛选合适的D-乳酸生产菌株和研究D-乳酸的发酵技术对拓宽D-乳酸的应用范围具有重要意义。      

高效转化木聚糖为乳酸植物乳杆菌L3的诱变选育及发酵条件的优化

以植物乳杆菌(Lactobacillus.pentosus)L3为出发菌株,进行紫外线和硫酸二乙酯(DES)诱变处理以提高其转化木聚糖生产乳酸的能力。经紫外线和DES诱变处理,得到突变菌株L.pentosusL35-6,其乳酸产量为71.2g/L,比诱变前提高了58.2%。并对其生长及乳酸发酵的条件进行了优化:最适温度为40℃,pH6.5,转速60r/min振荡培养发酵72h效果最好。     

Rhizopus oryzae LS1利用丢糟水解液发酵生产乳酸

研究了一株诱变米根霉Rhizopus oryzae LS-1利用丢糟水解液发酵生产乳酸的可行性。首先考察R.oryzae LS-1利用葡萄糖和木糖的糖代谢差异特性,并通过正交实验优化丢糟水解液发酵生产乳酸的工艺参数。结果表明,R.oryzae LS-1能代谢利用葡萄糖和木糖,且二者存在协同互补作用,有利于乳酸生成和糖酸转化,可用于木质纤维原料的乳酸生产。丢糟水解液发酵生产乳酸的实验表明,氯化铵是R.oryzae LS-1适宜的氮源。在接种量为3.0%、p H为6.5、发酵时间为96h、Ca CO3添加量为80g/L的条件下,乳酸生成浓度为13.27g/L,糖利用率为79.61%。说明诱变菌株R.oryzae LS-1具备发酵丢糟水解液制备乳酸的潜力。      

戊糖乳杆菌素pentocin LPEM818的初步纯化及特性研究

对戊糖乳杆菌(Lactobacillus pentosus)LPEM818所产戊糖乳杆菌素(pentocin)LPEM818进行了初步纯化,并对其生物学特性进行了研究。结果表明,采用80%硫酸铵沉淀和Sephadex G-25凝胶层析分离纯化后,细菌素的纯化倍数为11.09倍,回收率为6.4%;该细菌素在pH 2.0～8.0条件下稳定,121℃加热15 min保留84.52%的抑菌活性;对胰蛋白酶和蛋白酶K敏感;该细菌素的作用方式为杀菌;对供试的部分革兰氏阳性菌(G+)和革兰氏阴性菌(G-)具有较强抑制作用,因其抑菌谱较广,对多数致病菌和食品腐败菌有较好抑菌作用,所以具有作为食品生物防腐剂的潜在应用价值。     

戊糖乳杆菌S1-4所产抑菌物质的生物学特性研究

以内蒙古东部地区酸菜中分离得到的具有产抑菌活性的戊糖乳杆菌S1-4为供试菌,以大肠杆菌（Escherichia coli）3301为指示菌,采用双层琼脂扩散法对其所产抑菌物质的生物学特性进行研究。结果表明,戊糖乳杆菌S1-4所产的抑菌物经木瓜蛋白酶处理后,其抑菌活性明显降低,但对胃蛋白酶和胰蛋白酶不敏感。该抑菌物质经热处理后,对大肠杆菌仍保持较强的抑菌活性,在p H3.5⁵.5条件下,其抑菌活性较稳定。该菌株的发酵上清液分别经部分化学试剂和紫外线照射处理后,仍保持较好的抑菌活性。      

戊糖乳杆菌群体感应信号肽AIP的纯化及鉴定

目的:对戊糖乳杆菌31-1的群体感应信号肽进行提取、纯化和鉴定。方法:经硫酸铵盐析、超滤、疏水层析、凝胶层析和RP-HPLC步骤纯化戊糖乳杆菌31-1的群体感应信号肽。用MALDI-TOF质谱测定纯化后的小肽AIP的分子质量,并用EDMAN降解法对其N端氨基酸测序。用组氨酸蛋白激酶抑制剂——氯氰碘柳胺验证其群体感应信号肽分子功能。结果:纯化的信号肽AIP的比活力达160 000 IU/m L,纯度提高了800倍。该小肽的分子质量为2 985.16 u。其N端前15个氨基酸序列为NH2-KSSAYSLQMGATAIK。氯氰碘柳胺所造成的戊糖乳杆菌31-1细菌素合成功能的缺失,不能通过添加小肽AIP而得以恢复,证实了小肽AIP的群体感应信号肽分子功能。结论:群体感应信号肽AIP的获得为戊糖乳杆菌31-1细菌素的发酵调控奠定了基础。     

Effect of simulated acidic and salty fermentation conditions on kinetic growth parameters and probiotic potential of Lactobacillus acidipiscis and Lactobacillus pentosus

The technological stress during food processing may alter the beneficial interactions of food-associated Lactobacillus on the host. The effect of initial combined acidic (pH 5.0) and osmotic stress (NaCl 5 %, w/v with or without CaCl₂ 0.02 %, w/v) typically present in vegetable fermentations and cheesemaking were investigated. The growth kinetics prediction, hydrophobicity, auto-aggregation, in vitro survival and adhesion were determined in Lactobacillus acidipiscis and Lactobacillus pentosus isolated from a Mexican salty cheese. The growth was modelled with the modified Gompertz equation (R > 0.99; RMSE < 0.01). The lag phase increased (>10 h), and auto-aggregation was reduced. Hydrophobicity and S-layer total proteins varied. Gut survival reduced >4 log 10 units compared to the initial intake. Nevertheless, adhesion remained the same. Hence, combined stress affected the technological and physiological properties of both strains negatively. Further research is needed to identify the effect of the combined stress on the potential bioactivity of the strains.     

红景天和轮叶党参混合提取物戊糖乳杆菌发酵条件及抗氧化作用分析

目的：优化红景天和轮叶党参混合提取物（mixed extracts from Rhodiola sachalinensis and Codonopsis lanceolata,RCME）戊糖乳杆菌发酵条件,分析发酵产物的主要活性成分组成与含量,评价其抗氧化作用。方法：RCME中接种戊糖乳杆菌,以发酵液中的pH值、总酸含量及生物量为指标,研究发酵所需的最佳时间、培养基组成（红景天和轮叶党参混合比例）、RCME最佳质量浓度;采用比色法测定RCME发酵前后的总蛋白质、中性糖、酸性多糖及总酚含量变化,采用高效液相色谱法分析RCME发酵前后的红景天苷和酪醇含量变化,采用1,1-二苯基-2-三硝基苯肼（1,1-diphenyl-2-picrylhydrazyl,DPPH）自由基和2,2’-联氮-二（3-乙基-苯并噻唑-6-磺酸）二铵盐自由基（2,2’-azino-bis (3-ethylbenzthiazoline-6-sulfonic acid) diammonium salt radical,ABTS＋·）清除实验评价RCME发酵物的抗氧化作用。结果：单纯红景天提取物发酵24～72?h,其pH值和总酸含量无明显变化,随着发酵时间延长,乳杆菌数明显减少（P＜0.01）。红景天和轮叶党参混合质量比达到3∶2,发酵24h后发酵液的pH值下降幅度和总酸含量升高的幅度最大,发酵48?h时后,pH值和总酸含量无明显的变化,乳杆菌数有所减少。可溶性固形物质量分数在1.0%～3.0%范围内,红景天苷转化为酪醇的转化率无明显差异。RCME发酵24?h后,红景天苷含量显著减少（P＜0.01）,酸性多糖、总酚、蛋白质及酪醇含量均显著增加（P＜0.05或P＜0.01）。RCME发酵后DPPH自由基和ABTS＋·清除能力明显增强。结论：RCME发酵条件为红景天-轮叶党参混合比例3∶2、可溶性固形物质量分数1.0%～3.0%、发酵时间24?h。     

嗜酸乳杆菌乳酸高产菌株的紫外诱变选育

通过紫外诱变提高嗜酸乳杆菌的产乳酸能力,通过正交试验确定了诱变参数：20W紫外灯,菌液稀释至OD600O．65左右,照射60s,照射距离为30．5cm,此条件的致死率为92．5％。诱变处理后的茵体产酸速率明显高于出发菌株,突变菌株在培养36h时产酸量达到最大值10．83g／L,而出发菌株最大值为10．41g／L,且突变菌株具有稳定的遗传性。     

戊糖乳杆菌S1-4产抑菌物质发酵条件的优化及其抑菌谱

通过单因素及正交试验对戊糖乳杆菌S1-4产抑菌物质的发酵条件进行了优化,并其对敏感菌株的抑菌效果进行了探讨。结果表明,戊糖乳杆菌S1-4产抑菌物质的最佳培养条件为：接种量2.0%,培养基初始pH 5.5,32 ℃培养26 h,其抑菌圈平均直径可达（30.08±0.69） mm,优化前其抑菌圈平均直径为（24.41±0.25）mm,优化后抑菌活性提高了0.23 倍。菌株S1-4所产抑菌物质对所试12 种G＋和G－致病菌的生长都起到抑制作用,呈现出较广谱的抑菌特性。因此,菌株S1-4具有可作为食品生物防腐剂的潜在应用价值。     

Lactobacillus kefiranofaciens发酵生产Kefiran多糖的初步研究

研究了发酵条件(温度、起始pH、接种量等)对乳酸菌Lactobacillus kefiranofaciens发酵产Kefiran多糖的影响,并对菌体的形态及发酵特性进行了研究。研究结果表明,菌体形态在生长期间会发生改变,为短杆状-链杆状-短杆状;胞外多糖是从菌体生长期开始合成的;接种量为10%、起始pH为5.3、培养温度为25℃时有利于多糖的合成;CaCO3、Tween80和NaCl的添加量分别为1.2%、4%和0.25%时,胞外多糖的产量可达1.50g/L。     

Direct starch conversion into L‐(+)‐lactic acid by a novel amylolytic strain of Lactobacillus paracasei B41

A new amylolytic strain of Lactobacillus paracasei able to convert starch directly into L ‐(+)‐lactic acid (LA) was isolated. The identification of the by 16S rDNA sequencing proved that this strain, B41, is the first amylolytic representative of Lactobacillus casei group. The amylase activity assay revealed that L. paracasei B41 produced extracellular amylolytic enzyme, reaching 62 U/mL in the cell‐free supernatants. The optimal conditions for its action were pH 5.0 and temperature 45°C. The gene amy1 (1779 bp) encoding the putative B41 amylopullulanase was cloned, sequenced, and analyzed. The deduced protein contained a leader peptide of 28 amino acids and a mature peptide of 564 amino acids. Compared to the amylases of closely related species, B41 enzyme had several amino acid substitutions. An inducible control at amy1 expression was demonstrated. The starch fermentation abilities of L. paracasei B41 were studied in batch processes performed with and without pH control. The highest amount of LA from starch was obtained during 48 h fermentation from 40 g/L substrate at pH maintained at 5.0–37.3 g/L. In addition, 93.3% starch conversion into LA and the highest reported productivity for 24 h were achieved – 1.30 g/L/h.     

植物乳杆菌生物合成苯乳酸条件优化研究

以苯丙酮酸为底物,以植物乳杆菌静息细胞为催化剂,对苯乳酸生物合成条件进行了优化。 试验首先确定了不同因素对苯 乳酸产量的影响,然后通过正交试验优化了最佳合成条件。 结果表明,细胞培养时间为18 h时催化活性最高,且在温度为35 ℃,pH 为6.5,静息细胞、苯丙酮酸和葡萄糖质量浓度分别为80 g/L、3.0 g/L和4.0 g/L的催化条件下,经过4 h转化,苯乳酸产量达到最高,为 1.68 g/L。 在最优条件下,静息细胞重复利用4次,苯乳酸产量没有显著降低,表现出了植物乳杆菌转化苯丙酮酸合成苯乳酸的良好稳 定性。     

Wheat dough rheology and bread quality effected by Lactobacillus brevis preferment,dry sourdough and lactic acid addition

The influence of chemical and biological acidification on dough rheological properties and bread quality has been investigated. Two different flour types were used. Dough was chemically acidified with lactic acid. Two types of biologically acidified dough were prepared: dough with dry sourdough and with a Lactobacillus brevis preferment. Wheat dough rheological properties were investigated using the Farinograph, Extensograph and Amylograph. The baking response was also determined using standard baking tests. Addition of acidifiers resulted in firmer doughs with less stability, decreased extensibility and decreased gelatinisation maximum. The biological acidifiers increased the bread specific volume. Lactic acid addition had no influence on bread specific volume. In general, biological and chemical acidification decreased bread hardness. The addition of dry sourdough significantly decreased the lightness and increased the yellowness and redness of the bread crumb. The crust chroma, hue angle and brownness index were significantly changed by addition of acidifiers.     

向日葵籽乳酸饮料的研制

以向日葵为籽原料，磨浆后经乳酸发酵制取乳酸饮料，并通过对其制造过程的研究以及感观和风味的探讨，最终确定了向日葵籽乳酸饮料的最佳工艺和配方。