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Investigation of lactic acid bacteria mediated bioprotection with applications in cereal industry. Case-study: malting process

机译:乳酸菌介导的生物保护作用及其在谷物工业中的应用。案例研究:制麦过程

摘要

Antifungal compounds produced by Lactic acid bacteria (LAB) metabolites can be natural and reliable alternative for reducing fungal infections pre- and post-harvest with a multitude of additional advantages for cereal-base products. Toxigenic and spoilage fungi are responsible for numerous diseases and economic losses. This thesis includes an overview of the impact fungi have on aspects of the cereal food chain. The applicability of LAB in plant protection and cereal industry is discussed in detail. Specific case studies include Fusarium head blight, and the impact of fungi in the malting and baking industry. The impact of Fusarium culmorum infected raw barley on the final malt quality was part of the investigation. In vitro infected barley grains were fully characterized. The study showed that the germinative energy of infected barley grains decreased by 45% and grains accumulated 199 μg.kg-1 of deoxynivalenol (DON). Barley grains were subsequently malted and fully characterized. Fungal biomass increased during all stages of malting. Infected malt accumulated 8-times its DON concentration during malting. Infected malt grains revealed extreme structural changes due to proteolytic, (hemi)-cellulolytic and starch degrading activity of the fungi, this led to increased friability and fragmentation. Infected grains also had higher protease and β-glucanase activities, lower amylase activity, a greater proportion of free amino and soluble nitrogen, and a lower β-glucan content. Malt loss was over 27% higher in infected malt when compared to the control. The protein compositional changes and respective enzymatic activity of infected barley and respective malt were characterized using a wide range of methods. F. culmorum infected barley grains showed an increase in proteolytic activity and protein extractability. Several metabolic proteins decreased and increased at different rates during infection and malting, showing a complex F. culmorum infection interdependence. In vitro F. culmorum infected malt was used to produce lager beer to investigate changes caused by the fungi during the brewing processes and their effect on beer quality attributes. It was found, that the wort containing infected malt had a lower pH, a higher FAN, higher β-glucan and a 45% increase in the purging rate, and led to premature yeast flocculation. The beer produced with infected malt (IB) had also a significantly different amino acid profile. IB flavour characterization revealed a higher concentration of esters, fusel alcohols, fatty acids, ketones, and dimethylsulfide, and in particular, acetaldehyde, when compared to the control. IB had a greater proportion of Strecker aldehydes and Maillard products contributing to an increased beer staling character. IB resulted in a 67% darker colour with a trend to better foam stability. It was also found that 78% of the accumulated mycotoxin deoxynivalenol in the malt was transferred into beer. A LAB cell-freesupernatant (cfs), produced in wort-base substrate, was investigated for its ability to inhibit Fusarium growth during malting. Wort was a suitable substrate for LAB exhibiting antifungal activity. Lactobacillus amylovorus DSM19280 inhibited 104 spores.mL-1 for 7 days, after 120 h of fermentation, while Lactobacillus reuteri R29 inhibited 105 spores.mL-1 for 7 days, after 48 h of fermentation. Both LAB cfs had significant different organic acid profiles. Acid-base antifungal compounds were identified and, phenyllactic, hydroxy-phenyllactic, and benzoic acids were present in higher concentrations when compared to the control. A 3 °P wort substrate inoculated With L. reuteri R29 (cfs) was applied in malting and successfully inhibited Fusarium growth by 23%, and mycotoxin DON by 80%. Malt attributes resulted in highly modified grains, lower pH, higher colouration, and higher extract yield. The implementation of selected LAB producing antifungal compounds can be used successfully in the malting process to reduce mould growth and mycotoxin production.
机译:乳酸菌(LAB)代谢产物产生的抗真菌化合物可以是减少收获前和收获后真菌感染的天然可靠替代品,对于谷物基产品具有许多其他优势。产毒真菌和腐败真菌是造成许多疾病和经济损失的原因。本文概述了真菌对谷物食物链方面的影响。详细讨论了乳酸菌在植物保护和谷物工业中的适用性。具体的案例研究包括镰刀菌枯萎病以及真菌在麦芽和烘焙行业中的影响。镰刀菌感染的大麦对最终麦芽品质的影响是研究的一部分。体外感染的大麦籽粒已得到充分表征。研究表明,被感染的大麦籽粒的发芽能降低了45%,谷物中积累了199μg.kg-1的脱氧雪腐烯醇(DON)。随后将大麦籽粒制成麦芽并充分表征。在制麦的所有阶段,真菌生物量均增加。麦芽过程中被感染的麦芽积累了其DON浓度的8倍。由于真菌的蛋白水解,(半)纤维素分解和淀粉降解活性,被感染的麦芽籽粒显示出极端的结构变化,这导致脆碎性增加和破碎。受感染的谷物还具有较高的蛋白酶和β-葡聚糖酶活性,较低的淀粉酶活性,较大比例的游离氨基和可溶性氮,以及较低的β-葡聚糖含量。与对照相比,受感染麦芽的麦芽损失高出27%以上。使用多种方法对受感染大麦和相应麦芽的蛋白质组成变化以及相应的酶活性进行了表征。细角镰刀菌感染的大麦籽粒显示出蛋白水解活性和蛋白质提取能力的增加。在感染和麦芽形成过程中,几种代谢蛋白以不同的速率下降和增加,显示出复杂的鳞茎镰刀菌感染相互依赖。用体外感染了F. culmorum的麦芽来生产大啤酒,以调查酿造过程中真菌引起的变化及其对啤酒品质属性的影响。发现含有麦芽汁的被感染麦芽的pH较低,FAN较高,β-葡聚糖较高,吹扫率提高了45%,导致酵母过早絮凝。用受感染的麦芽(IB)生产的啤酒也具有明显不同的氨基酸谱。 IB风味特征显示与对照相比,酯,杂醇,脂肪酸,酮和二甲硫特别是乙醛的浓度更高。 IB的Strecker醛和美拉德产品的比例更高,这有助于增加啤酒的陈旧性。 IB导致颜色变深67%,具有更好的泡沫稳定性的趋势。还发现麦芽中积累的霉菌毒素脱氧雪茄烯醇的78%已转移到啤酒中。研究了在麦芽汁基质中产生的无LAB细胞上清液(cfs)抑制麦芽过程中镰刀菌生长的能力。麦芽汁是用于LAB的具有抗真菌活性的合适底物。发酵120小时后,淀粉乳杆菌DSM19280抑制104个孢子。mL-17天,而发酵48小时后,罗伊氏乳杆菌R29抑制105个孢子。mL-17天。两种LAB cfs均具有明显不同的有机酸谱。鉴定了酸基抗真菌化合物,与对照相比,苯乳酸,羟基苯乳酸和苯甲酸的浓度更高。将接种罗伊氏乳杆菌R29(cfs)的3°P麦芽汁底物用于制麦芽,成功抑制镰刀菌生长23%,抑制真菌毒素DON 80%。麦芽的特性导致谷物高度改性,较低的pH,较高的着色度和较高的提取物收率。选定的LAB产生的抗真菌化合物的实施可成功用于麦芽制造过程中,以减少霉菌的生长和霉菌毒素的产生。

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