乳酸菌β-甘露聚糖酶研究进展

β-甘露聚糖酶属于糖苷水解酶（glycoside hydrolases,GH）家族,广泛存在于植物、动物和微生物中,主要水解产物为甘露低聚糖（mannan-oligosaccharides,MOS）。微生物来源的甘露聚糖酶种类多且活性高,但大部分安全性低,其中乳酸菌（lactic acid bacteria,LAB）甘露聚糖酶具有纯化步骤少、易于提取、安全性高等优点,能够满足食品级工业生产的需求,因此在保健品、饲料、饮料生产等工业中有重要应用前景。本文介绍了天然LAB甘露聚糖酶的产酶条件、分离纯化和酶学特性,简述了近年来甘露聚糖酶的LAB重组表达及其在果汁澄清和制备益生元MOS方面的应用,为LAB甘露聚糖酶的基础研究和开发提供参考和依据。     

Production of Bioactive Peptides from Lactic Acid Bacteria: A Sustainable Approach for Healthier Foods

Traditional fermented foods where lactic acid bacteria (LAB) are present have been associated with beneficial effects on human health, and some of those benefits are related to protein‐derived products. Peptides produced by LAB have attracted the interest of food industries because of their diverse applications. These peptides include ribosomally produced (bacteriocins) and protein hydrolysates by‐products (bioactive peptides), which can participate as natural preservatives and nutraceuticals, respectively. It is essential to understand the biochemical pathways and the effect of growth conditions for the production of bioactive peptides and bacteriocins by LAB, in order to suggest strategies for optimization. LAB is an important food‐grade expression system that can be used in the simultaneous production of peptide‐based products for the food, animal, cosmetic, and pharmaceutical industries. This review describes the multifunctional proteinaceous compounds generated by LAB metabolism and discusses a strategy to use a single‐step production process, using an alternative protein‐based media. This strategy will provide economic advantages in fermentation processes and will also provide an environmental alternative to industrial waste valorization. New technologies that can be used to improve production and bioactivity of LAB‐derived peptides are also analyzed.     

Invited review: Bioactive compounds produced during cheese ripening and health effects associated with aged cheese consumption

Traditionally, cheese is manufactured by converting fluid milk to a semisolid mass through the use of a coagulating agent, such as rennet, acid, heat plus acid, or a combination thereof. Cheese can vary widely in its characteristics, including color, aroma, texture, flavor, and firmness, which can generally be attributed to the production technology, source of the milk, moisture content, and length of aging, in addition to the presence of specific molds, yeast, and bacteria. Among the most important bacteria, lactic acid bacteria (LAB) play a critical role during the cheese-making process. In general, LAB contain cell-envelope proteinases that contribute to the proteolysis of cheese proteins, breaking them down into oligopeptides that can be subsequently taken up by cells via specific peptide transport systems or further degraded into shorter peptides and amino acids through the collaborative action of various intracellular peptidases. Such peptides, amino acids, and their derivatives contribute to the development of texture and flavor in the final cheese. In vitro and in vivo assays have demonstrated that specific sequences of released peptides exhibit biological properties including antioxidant, antimicrobial, anti-inflammatory, immunomodulatory, and analgesic/opioid activity, in addition to angiotensin-converting enzyme inhibition and antiproliferative activity. Some LAB also produce functional lipids (e.g., conjugated linoleic acid) with anti-inflammatory and anticarcinogenic activity, synthesize vitamins and antimicrobial peptides (bacteriocins), or release γ-aminobutyric acid, a nonprotein amino acid that participates in physiological functions, such as neurotransmission and hypotension induction, with diuretic effects. This review provides an overview of the main bioactive components present or released during the ripening process of different types of cheese.     

The Use and Effects of Lactic Acid Bacteria in Malting and Brewing with Their Relationships to Antifungal Activity,Mycotoxins and Gushing: A Review

Several metabolic properties of lactic acid bacteria (LAB) serve special functions, which directly or indirectly have impact on processes such as improved quality and safety and flavour development in the malting and brewing industry. LAB are widely distributed in nature and in spontaneous fermentations, often they are found to be the dominating microflora resulting in both the inhibition of spoilage bacteria and organisms. This review describes the applications of LAB in malting and brewing. Mycotoxins are naturally occurring toxic secondary metabolites of fungi that may be present in cereals. Several of these mycotoxins have been associated with human and animal diseases and are known to survive the brewing process. LAB have been shown to restrict the growth of the most important toxigenic fungi thereby reducing the formation of these harmful toxins. The occurrence of mycotoxins in cereals is discussed and their effect in beer is reviewed. The main features of this review are: (I) LAB starter cultures in malting and brewing (II) production of acid malt; (III) biological acidification of mash and wort in brewing; (IV) bacteriocins produced by LAB in brewing; (V) LAB and anti‐fungal activity; (VI) mycotoxins in cereals.     

Milk fat globule membrane glycoproteins: Valuable ingredients for lactic acid bacteria encapsulation?

The membrane (Milk Fat Globule Membrane – MFGM) surrounding the milk fat globule is becoming increasingly studied for its use in food applications due to proven nutritional and technological properties. This review focuses first on current researches which have been led on the MFGM structure and composition and also on laboratory and industrial purification and isolation methods developed in the last few years. The nutritional, health benefits and techno-functional properties of the MFGM are then discussed. Finally, new techno-functional opportunities of MFGM glycoproteins as a possible ingredient for Lactic Acid Bacteria (LAB) encapsulation are detailed. The ability of MFGM to form liposomes entrapping bioactive compounds has been already demonstrated. One drawback is that liposomes are too small to be used for bacteria encapsulation. For the first time, this review points out the numerous advantages to use MFGM glycoproteins as a protecting, encapsulating matrix for bacteria and especially for LAB.     

淫羊藿叶多糖工艺优化及抗氧化活性

探索和优化超声复合酶解提取淫羊藿叶粗多糖工艺。在单因素试验的基础上,通过正交试验优化复合酶添加量,再采用Box-Behnken设计和响应面优化超声复合酶解提取工艺参数。结果显示:不同酶添加量对多糖提取得率的影响次序是:纤维素酶果胶酶木瓜蛋白酶α-淀粉酶,最佳复合酶(木瓜蛋白酶、果胶酶、纤维素酶和α-淀粉酶)的添加量分别为50、250、200、100 U/g;最佳提取条件为提取温度46.8℃、超声时间42.3 min、p H 4.3、超声功率311 W。在此实验条件下,粗多糖提取得率为5.98%,与模型预测值(6.2%)接近。粗多糖通过琼脂糖离子交换和葡聚糖分子筛凝胶柱色谱分离纯化,得到3个主要多糖组分(EPs-1、EPs-2、EPs-3),多糖组分采用DPPH自由基、羟自由基、超氧阴离子自由基清除和铁离子还原能力实验进行了抗氧化活性评价。结果表明,超声复合酶提取作为一个高效和环保的提取技术,可以应用于从植物原料中提取活性成分;抗氧化活性实验显示3个多糖组分都具有显著的抗氧化活性,其活性呈添加量依赖关系。这些结果说明淫羊藿多糖可以探索作为潜在的抗氧剂应用于功能食品和药品。     

微生物脂肪酶的研究与应用

脂肪酶是一类能够催化酯的水解反应以及在非水相体系中催化脂肪酸和醇类发生酯化反应的酶类。随着酶学技术的快速发展,微生物脂肪酶也受到了越来越多的关注。作为生物催化剂,脂肪酶一直以来都是生物技术领域中最重要的一类酶。本文探讨了脂肪酶的来源、理化性质、脂肪酶活力测定,同时对脂肪酶的非水相催化特性以及脂肪酶在食品工业,医药、洗涤剂、皮革、造纸和生物柴油工业领域中的应用进行了讨论。     

FUNDAMENTALS AND PERSPECTIVES FOR THE USE OF BACTERIOCINS PRODUCED BY LACTIC ACID BACTERIA IN MEAT PRODUCTS

Bacteriocins of lactic acid bacteria (LAB) are proteinaceous compounds that may present antimicrobial activity towards important foodborne pathogens and spoilage-related microflora. Due to these properties, bacteriocin-producing strains or purified bacteriocins have a great potential of use in biologically based food preservation systems. Despite the growing number of articles describing the isolation of bacteriocinogenic strains, genetic determinants for production, as well as the purification and biochemical characterization of these inhibitory substances, there are only limited reports of successful application of bacteriocins to meats.

This paper presents a critical review of the methods available for screening of bacteriocin-producing LAB strains from meats and also discusses the proposed mechanisms of action for LAB bacteriocins. Additionally, an overview of the Brazilian experience in the application of LAB bacteriocins to meats and meat products is given.     

FUNDAMENTALS AND PERSPECTIVES FOR THE USE OF BACTERIOCINS PRODUCED BY LACTIC ACID BACTERIA IN MEAT PRODUCTS

Bacteriocins of lactic acid bacteria (LAB) are proteinaceous compounds that may present antimicrobial activity towards important foodborne pathogens and spoilage-related microflora. Due to these properties, bacteriocin-producing strains or purified bacteriocins have a great potential of use in biologically based food preservation systems. Despite the growing number of articles describing the isolation of bacteriocinogenic strains, genetic determinants for production, as well as the purification and biochemical characterization of these inhibitory substances, there are only limited reports of successful application of bacteriocins to meats.

This paper presents a critical review of the methods available for screening of bacteriocin-producing LAB strains from meats and also discusses the proposed mechanisms of action for LAB bacteriocins. Additionally, an overview of the Brazilian experience in the application of LAB bacteriocins to meats and meat products is given.     

乳酸菌食品级质粒及应用的研究进展

乳酸菌是食品发酵工业中重要的益生菌,利用分子生物学技术构建的菌种对食品产业发展及人类健康具有重要影响。本文主要通过介绍乳酸菌食品级系统的基本要求、食品级质粒的元件组成、食品级质粒构建策略及其应用的研究进展,展示了乳酸菌食品级分子操作系统的建立对乳酸菌的深层次开发利用所具有的重要意义。     

Valorization of fish byproducts: Sources to end-product applications of bioactive protein hydrolysate

Fish processing industries result in an ample number of protein-rich byproducts, which have been used to produce protein hydrolysate (PH) for human consumption. Chemical, microbial, and enzymatic hydrolysis processes have been implemented for the production of fish PH (FPH) from diverse types of fish processing byproducts. FPH has been reported to possess bioactive active peptides known to exhibit various biological activities such as antioxidant, antimicrobial, angiotensin-I converting enzyme inhibition, calcium-binding ability, dipeptidyl peptidase-IV inhibition, immunomodulation, and antiproliferative activity, which are discussed comprehensively in this review. Appropriate conditions for the hydrolysis process (e.g., type and concentration of enzymes, time, and temperature) play an important role in achieving the desired level of hydrolysis, thus affecting the functional and bioactive properties and stability of FPH. This review provides an in-depth and comprehensive discussion on the sources, process parameters, purification as well as functional and bioactive properties of FPHs. The most recent research findings on the impact of production parameters, bitterness of peptide, storage, and food processing conditions on functional properties and stability of FPH were also reported. More importantly, the recent studies on biological activities of FPH and in vivo health benefits were discussed with the possible mechanism of action. Furthermore, FPH-polyphenol conjugate, encapsulation, and digestive stability of FPH were discussed in terms of their potential to be utilized as a nutraceutical ingredient. Last but not the least, various industrial applications of FPH and the fate of FPH in terms of limitations, hurdles, future research directions, and challenges have been addressed.     

乳酸菌细菌素研究进展及其在水产养殖和加工中的应用

乳酸菌细菌素是一种由乳酸菌在核糖体内合成、具有抑菌活性的多肽或蛋白质。由于乳酸菌通常被认为是安全的微生物,因此由它产生的细菌素受到了广泛地关注。目前由于大部分乳酸菌细菌素抑菌机理研究还不够深入,在一定程度上限制了它的发展和应用。本文对现有乳酸菌细菌素研究成果进行总结,并对细菌素进行了系统分类,阐述了各类乳酸菌细菌素对革兰氏阳性菌的作用机理,最后介绍了乳酸菌细菌素在水产养殖、水产品加工贮藏过程中的应用,旨在为乳酸菌细菌素的应用提供新的探索和基础理论依据。     

Probiotics and Its Functionally Valuable Products—A Review

During the past two decades probiotic bacteria have been increasingly proposed as health promoting bacteria in variety of food system, because of its safety, functional, and technological characteristics. Commonly, Lactobacillus spp., Bifidobacterium spp., Saccharomyces boulardii, and some other microorganisms have been considered as probiotic strains. Possibly these bacterial strains exerted several beneficial effects into gastrointestinal tract of host while administered with variety of food system. Lactic acid bacteria (LAB) usually produce antimicrobial substances like bacteriocin which have broad spectrum of antagonist effect against closely related Gram positive and Gram negative pathogens. LAB strains often produce polymeric substances such as exopolysaccharides (EPS) which increase the colonization of probiotic bacteria by cell–cell interactions in gastrointestinal tract. LAB also produces biosurfactant which showed that the wide range of antimicrobial activity against bacterial pathogen as well as its antiadhesive properties reduces the adhesion of pathogens into gastric wall membrane. Furthermore, LAB strains have also been reported for production of antioxidants which are ability to scavenge the free radicals such as superoxide anions and hydroxyl radicals. For this sense, this review article is mainly focused on the ecology, biosynthesis, genetics, target sites, and applications of bacteriocins and EPS from LAB strains. Moreover, this review discusses about the production and functions of nutritive essential element folate and iron chelating agent such as siderophores from LAB.     

Esterases of lactic acid bacteria and cheese flavour: Milk fat hydrolysis,alcoholysis and esterification

Free fatty acids (FFAs) and esters derived from FFAs are important flavour compounds in cheese. Evidence is provided that esterases of lactic acid bacteria (LAB) catalyse not only hydrolysis of milk fat glycerides to release FFAs, but also synthesis of esters from glycerides and alcohols via a transferase reaction. The esterases of LAB prefer di- and monoglycerides for both hydrolysis and ester synthesis and are, in fact, alcohol acyltransferases that use both water (hydrolysis) and alcohol (alcoholysis) as acyl acceptors. Therefore, esterases of LAB can impact on both the lipolytic and ester flavours of cheese. The impact of esterases of LAB on cheese flavour can be controlled by manipulating the amount of the esterase, by regulating alcohol availability and/or by increasing the mono- and diglyceride composition of milk fat. In addition, esterification may play a role in ester synthesis in hard cheeses (e.g. Italian type) where water activity is low.     

Release and partial characterization of cell-envelope proteinases fromLactococcus lactis subsp.lactis IFPL 359 andLactobacillus casei subsp. casei IFPL 731 isolated from raw goat,s-milk cheese

Different methods of releasing the cell-envelope proteinase (CEP) fromLactococcus lactis IFPL 359 (Lc-CEP) andLactobacillus casei IFPL 731 (Lb-CEP) have been tested. Release of Lc-CEP was higher in Ca^2+-free buffer than in the presence of lysozyme and Ca^2+-. Lb-CEP was not soluble in Ca^2+--free buffer, making necessary the use of chelating agents such as ethylenediaminetetraacetate (EDTA) to attain release yields of 15–20%. Solubilizing the cell wall oflb.casei using lysozyme and mutanolysin improved CEP release yields, even in the presence of Ca^2+-. Two differently charged chromophoric peptides were degraded by whole cells and the soluble fractions studied at different hydrolysis rates in both the strains considered. Based on the specificity of these CEPs for the different substrates, the two proteinases can be placed in the same class as the CEP_I/III mixed-type variants that have been identified in lactococcal proteinases. In both strains ß-casein was hydrolysed more rapidly thanα _s-cascin.     

Applications of sample preparation techniques in the analysis of pesticides and PCBs in food

Pesticides and polychlorinated biphenyls (PCBs) are found in various parts of the environment in quite small concentrations, but they accumulate and thus become a threat to human health and life. A review is focused on the application of some popular techniques for sample preparation in analysis of these compounds in food. Even with the emergence of advanced techniques of final analysis, complex matrices, such as food, require extensive sample extraction and purification. Traditional sample preparation techniques are time consuming and require large amount of solvents, which are expensive, generate considerable waste, contaminate the sample and can enrich it for analytes. There have been many sample preparation techniques proposed to meet the requirements connected with the multiplicity of food. Optimal sample preparation can reduce analysis time, sources of error, enhance sensitivity and enable unequivocal identification and quantification. Sample extraction and purification techniques are discussed and their most recent applications in food analysis are provided. This review pointed out that sample preparation is the critical step.     

龙牙楤木中酚类物质提取方法的比较

为筛选适合龙牙楤木中酚类物质的提取方法。本文以龙牙楤木嫩叶为原料,采用溶剂浸提法、溶剂回流法、超声辅助法、微波辅助法以及复合酶法提取龙牙楤木嫩叶中酚类物质,对酚类物质的得率进行比较分析,筛选出最佳提取方法,并在其单因素基础上,进一步利用正交试验优化各工艺参数。结果表明,五种提取方法中复合酶法提取酚类物质效果最好;正交试验得到复合酶法提取的最佳工艺参数为:复合酶（果胶酶和各种碳水化合物酶组成）用量120 μL/g（酶量/样品量）,酶解温度50 ℃,酶解pH6,酶解时间1.5 h,乙醇体积分数50%,此条件下龙牙楤木中总酚得率为7.24%,总黄酮得率为3.17%。通过正交实验优化复合酶法提取龙牙楤木嫩叶中酚类物质,在最佳工艺参数下提取的效果理想,可用于后续对酚类物质纯化、鉴定等研究。     

Original features of cell-envelope proteinases of Lactobacillus helveticus. A review

Lactobacillus helveticus is a lactic acid bacterium very used in fermented milks and cheese. The rapid growth of L. helveticus in milk is supported by an efficient cell envelope proteinase (CEP) activity, due to subtilisin-like serine proteases. These enzymes play also crucial roles in texture and flavor formation in dairy products as well as in generating in situ bioactive peptides. In L. helveticus, several genes encoding putative CEPs were detected and characterized by a large intraspecific diversity; little is known about regulation of expression of CEP-encoding genes. Anchored at the bacterial surface, CEPs are large-sized enzymes (> 150 kDa) hydrolyzing β- and α_s1-casein as well. Substrate cleavages occur after almost all types of amino acids residues, but mass spectrometry analysis revealed L. helveticus strains with specific profiles of substrate hydrolysis, which could explain identification of strains associated with interesting technological properties. In this review, the most recent data regarding CEP-encoding genes, CEP activities toward caseins and L. helveticus strain diversity are discussed.     

Saponins: properties, applications and processing

Saponins are a diverse group of compounds widely distributed in the plant kingdom, which are characterized by their structure containing a triterpene or steroid aglycone and one or more sugar chains. Consumer demand for natural products coupled with their physicochemical (surfactant) properties and mounting evidence on their biological activity (such as anticancer and anticholesterol activity) has led to the emergence of saponins as commercially significant compounds with expanding applications in food, cosmetics, and pharmaceutical sectors. The realization of their full commercial potential requires development of new processes/processing strategies to address the processing challenges posed by their complex nature. This review provides an update on the sources, properties, and applications of saponins with special focus on their extraction and purification. Also reviewed is the recent literature on the effect of processing on saponin structure/properties and the extraction and purification of sapogenins.