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Phenolic profile, total bioactive contents, and antioxidant activity of pear fruits 梨果的酚类概况、生物活性总含量和抗氧化活性
Pub Date : 2024-07-20 DOI: 10.1016/j.focha.2024.100780

This work describes the profiling of phenolic compounds, total phenolic contents, total flavonoid contents, total anthocyanins, and radical scavenging activity of two important cultivars of pears grown in Pakistan. The HPLC-DAD reversed phase chromatography was used to study phenolic compounds. The spectrophotometry was done to measure the total flavonoid contents, total phenolic contents, total anthocyanin contents, and radical scavenging activity. Results showed 21 phenolic compounds in the extract of Comice pears and 16 in Bosc pears. These include 3-caffeoylquinic acid, p-coumaroylhexose-4-hexoside, 5-coumaroylquinic acid, cis-5-coumaroylquinic acid, (+)-catechin-3-glucoside, ellagic acid dihexoside, quercetin-3-rutinoside, eriodictyol-7-rutinoside, 3-hydroxyphloretin-6′-hexoside, (−)-epigallocatechin, quercetin pentosyl hexoside, (−)-epigallocatechin-3-gallate, epicatechin-3-(4-methyl)-gallate, rosmarinic acid, cyanidin-3,5-diglucoside, 4-caffeoyl-5-coumaroylquinic acid were major identified phenolic compounds in both Comice and Bosc pears. Except for total anthocyanin contents, there were significant differences between the selected cultivars in terms of total phenolic contents, total flavonoid contents, and radical scavenging activity values. In conclusion, the selected pear samples contain important phenolic compounds and possess antioxidant properties.

本研究描述了巴基斯坦种植的两个重要梨品种的酚类化合物、总酚含量、总黄酮含量、总花青素含量和自由基清除活性。采用 HPLC-DAD 反相色谱法研究酚类化合物。分光光度法测量了总黄酮含量、总酚含量、总花青素含量和自由基清除活性。结果表明,康美斯梨提取物中含有 21 种酚类化合物,博斯克梨提取物中含有 16 种酚类化合物。其中包括 3-咖啡酰奎宁酸、对香豆酰己糖-4-己糖苷、5-香豆酰奎宁酸、顺式-5-香豆酰奎宁酸、(+)-儿茶素-3-葡萄糖苷、鞣花酸二己糖苷、槲皮素-3-芸香糖苷、二碘酪醇-7-芸香糖苷、3-羟基香紫苏-6′-己糖苷、(-)-表没食子儿茶素、槲皮素五糖基六糖苷、(-)-表没食子儿茶素-3-没食子酸酯、表儿茶素-3-(4-甲基)没食子酸酯、迷迭香酸、青花素-3,5-二葡萄糖苷、4-咖啡酰-5-香豆酰奎宁酸是在康美斯梨和博斯克梨中鉴定出的主要酚类化合物。除总花青素含量外,所选栽培品种之间在总酚含量、总黄酮含量和自由基清除活性值方面存在显著差异。总之,所选梨样品含有重要的酚类化合物,具有抗氧化特性。
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引用次数: 0
Effect of distinct drying approach on bioactive compounds and nutritional profiling of Ulva intestinalis and Padina tetrastromatica from the Bay of Bengal, Bangladesh 不同干燥方法对孟加拉国孟加拉湾的肠莼和四鳃莼的生物活性化合物和营养成分的影响
Pub Date : 2024-07-19 DOI: 10.1016/j.focha.2024.100778

Marine algae have been visualized as fostering a prevalent actuation of bioactive substances that have noteworthy therapeutic effects. The current investigations deployed a trio of separate drying methods, i.e., oven drying, freeze drying, and sun drying, to quantify the total phenolic content (TPC), total flavonoid content (TFC), and dietary vitality of Ulva intestinalis and Padina tetrastromatica. By mining and scrutinizing it to the quercetin and gallic acid standards, TPC and TFC were derived. Among three drying techniques, freeze drying asserted higher TPC (12.59 ± 1.07 in U. intestinalis and 68.74 ± 1.95 mg of GA/g in P. tetrastromatica) and TFC (7.92 ± 0.97 in U. intestinalis and 58.15 ± 2.05 mg of quercetin/g in P. tetrastromatica) values. Ulva intestinalis and P. tetrastromatica also contain a high amount of protein (18.34% and 8.7%, respectively). The findings demonstrated that green seaweed possessed higher levels of potassium (2.20%) and molybdenum (2.41%) and brown seaweed showed higher amounts of potassium (2.41%) and calcium (1.11%). Where rapid drying methods are recommended to enhance constituent levels, freeze-drying could be an attractive alternative to oven and sun drying, as it addresses the drawbacks associated with prolonged drying periods.

海藻被认为普遍含有生物活性物质,具有显著的治疗效果。目前的研究采用了三种不同的干燥方法,即烘箱干燥、冷冻干燥和日光干燥,以量化肠道莼菜和四色莼菜的总酚含量(TPC)、总黄酮含量(TFC)和膳食活力。通过提取并与槲皮素和没食子酸标准进行比对,得出了总酚含量和总黄酮含量。在三种干燥技术中,冷冻干燥的 TPC 值(肠莼为 12.59 ± 1.07,没食子酸为 68.74 ± 1.95 毫克/克)和 TFC 值(肠莼为 7.92 ± 0.97,没食子酸为 58.15 ± 2.05 毫克槲皮素/克)较高。此外,肠莼和四色鲈还含有大量蛋白质(分别为 18.34% 和 8.7%)。研究结果表明,绿色海藻的钾含量(2.20%)和钼含量(2.41%)较高,而褐色海藻的钾含量(2.41%)和钙含量(1.11%)较高。在建议采用快速干燥方法提高成分含量的情况下,冷冻干燥可能是烤箱干燥和日光干燥的一种有吸引力的替代方法,因为它解决了与长时间干燥相关的缺点。
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引用次数: 0
Development of functional probiotic yogurt from buffalo milk supplemented with red beetroot (Beta vulgaris L.) as an antioxidant, natural colorant, and starter growth stimulant 利用水牛奶开发功能性益生菌酸奶,并辅以红甜菜根(Beta vulgaris L.)作为抗氧化剂、天然着色剂和启动生长刺激剂
Pub Date : 2024-07-19 DOI: 10.1016/j.focha.2024.100776

The physicochemical, sensorial, and probiotics viability properties of supplemented probiotic yogurt with beet juice (BJ) at various ratios (0–5)% in fresh and stored yogurts were determined. 1,1-diphenyl-2-picrylhydrazyl (DPPH) scavenging activity (%) and minerals content of BJ-yogurts were increased. The color aspects (ΔE) were improved with the increasing of BJ. BJ-yogurts at 1 and 3 % were color-stable when stored in the refrigerator (5 ± 1 °C) for 9 days. The syneresis reached of 42.9, 43.0, 43.5, and 43.5 mL for control, 1 %, 3 %, and 5 % BJ, respectively in stored yogurts for 12 days. Moreover, the probiotic yogurt viability in BJ-yogurts treatments was found to be higher (p < 0.05) than control which recorded of 7.0 ± 0.08, 7.22±0.04, 7.34±0.002 log cfu/mL for streptococci, lactobacilli, and bifidobacteria counts respectively in 5 % BJ treatment, on the other hand, in control recorded of 6.76±0.07, 7.11±0.06, 6.66±0.030 log cfu/mL for streptococci, lactobacilli, and bifidobacteria counts respectively; indicating that BJ had a stimulating effect on the additive starter. Fresh yogurts with 1 % BJ had superior sensorial scores than other treatments, and generally maintained desirable organoleptic attributes for up to 9 days. This study provides a nutritious, low-cost, stable, and attractive colored yogurts from buffalo milk without negative impact on physiochemical qualities.

测定了在新鲜和储存的酸奶中以不同比例(0-5)%添加甜菜汁(BJ)的益生菌酸奶的理化、感官和益生菌活力特性。BJ 酸奶的 1,1-二苯基-2-苦基肼(DPPH)清除活性(%)和矿物质含量均有所提高。色度(ΔE)随 BJ 的增加而提高。1 % 和 3 % 的 BJ 酸奶在冰箱(5 ± 1 °C)中储存 9 天后颜色稳定。在存放 12 天的酸奶中,对照组、1%、3% 和 5% 的 BJ 酸奶的滞留率分别为 42.9、43.0、43.5 和 43.5 mL。此外,还发现 BJ 酸奶中益生菌的存活率比对照组高(p < 0.05),在 5 % BJ 处理中,链球菌、乳酸菌和双歧杆菌的存活率分别为 7.0 ± 0.08、7.22±0.04、7.34±0.002 log cfu/mL,而在对照组中,链球菌、乳酸菌和双歧杆菌的存活率分别为 6.76±0.07、7.22±0.04、7.34±0.002 log cfu/mL。76±0.07、7.11±0.06、6.66±0.030 log cfu/mL。添加了 1 % BJ 的新鲜酸奶的感官评分优于其他处理,而且在长达 9 天的时间里一般都能保持理想的感官特性。这项研究提供了一种营养丰富、成本低廉、稳定且美观的水牛奶彩色酸奶,且不会对理化品质产生负面影响。
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引用次数: 0
Fortification of bread with mango peel and pulp as a source of bioactive compounds: A comparison with plain bread 用芒果皮和果肉作为生物活性化合物来源强化面包:与普通面包的比较
Pub Date : 2024-07-18 DOI: 10.1016/j.focha.2024.100783

In this study, fortified bread was prepared with the incorporation of mango peel powder (MPP), and mango pulp powder (MPuP). Different concentrations of MPP (2.5 %, 5.0 %, and 7.5 %) and MPuP (7.5 %) were added to improve the functional and bioactive properties of bread. Pieces of bread incorporated with MPP demonstrated a significant improvement in bioactive properties over control and MPuP-fortified bread, where total phenol content elevated from 5.08 ± 0.99 to 54.29 ± 2.34 mM GAE/100 g DM and total flavonoid content raised from 5.7 ± 0.45 to 50.72 ± 3.40 mM QE/100 g DM. The inclusion of MPP increased antioxidant activity, with a range of 0.585 ± 0.12 to 18.20 ± 0.87 mM TE/100 g DM in DPPH assay and 10.75 ± 0.98 to 30.24 ± 2.12 μM Fe (II) E/g DM in FRAP assay, at the highest (7.5 %) level of MPP incorporation. However, the specific volume and weight loss percentage of breads were decreased with the incorporation of MPP. The whiteness index (WI) dropped, and the brownness index (BI) improved with the degree of MPP integration. Bread volume and aroma drop considerably at the level of 7.5 % MPP incorporation, however, a slight mango flavor was noticed with the incorporation of 7.5 % MPuP in breads. Thus, 5 % MPP inclusion in wheat bread was deemed an optimum amount without compromising the sensory and physical characteristics along with national properties.

本研究制备了添加芒果皮粉(MPP)和芒果果肉粉(MPuP)的强化面包。为了改善面包的功能性和生物活性,面包中添加了不同浓度的芒果皮粉(2.5%、5.0% 和 7.5%)和芒果果肉粉(7.5%)。与对照组和添加 MPuP 的面包相比,添加了 MPP 的面包在生物活性特性方面有显著改善,总酚含量从 5.08 ± 0.99 mM GAE/100 g DM 提高到 54.29 ± 2.34 mM GAE/100 g DM,总黄酮含量从 5.7 ± 0.45 mM QE/100 g DM 提高到 50.72 ± 3.40 mM QE/100 g DM。添加 MPP 提高了抗氧化活性,在 DPPH 试验中,抗氧化活性范围为 0.585 ± 0.12 至 18.20 ± 0.87 mM TE/100 g DM,在 FRAP 试验中,抗氧化活性范围为 10.75 ± 0.98 至 30.24 ± 2.12 μM Fe (II) E/g DM,MPP 添加量最高(7.5%)。然而,面包的比容和失重率随着 MPP 的添加而降低。白度指数(WI)随着 MPP 添加量的增加而下降,褐度指数(BI)则随着添加量的增加而提高。面包的体积和香味在 MPP 含量为 7.5 % 时明显下降,但在面包中添加 7.5 % 的 MPuP 时,会有轻微的芒果香味。因此,在小麦面包中添加 5 % 的多氯联苯酚被认为是不影响感官和物理特性以及国民特性的最佳添加量。
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引用次数: 0
Dense phase carbon dioxide (DPCD) inactivation of microorganisms and enzymes, and its application in food: A review 密相二氧化碳(DPCD)灭活微生物和酶及其在食品中的应用:综述
Pub Date : 2024-07-17 DOI: 10.1016/j.focha.2024.100782

A surge of interest in using non-thermal, non-toxic, and chemical-free food preservation methods is seen in recent years. At the same time, there is a requirement for fresh, minimally processed, and nutritious food in the market. Dense phase carbon dioxide (DPCD) is a non-thermal method of inactivating enzymes and microorganisms at temperatures below standard pasteurization to avoid thermal effects. Diverse research has been conducted to understate the mechanism and its effect on microorganisms such as bacteria, yeast, molds, and enzymes throughout the last few decades. The effects of this technology have been promising in the case of many liquid foods. Furthermore, researchers are working on the effect of DPCD in combination with other non-thermal processes such as irradiation, PEF, ultrasound, and HPP. This review compiles a substantial quantity of research on DPCD and provides an overview of DPCD processing equipment, mechanism, applications in liquid and solid foods, the emerging interest of DPCD in hurdle technology, and highlights its efficiency against microorganisms and enzymes, as well as its application in the food processing industry. DPCD offers new opportunities for better microbe control, enzyme inactivation at softer process settings, and improved quality and nutrient retention.

近年来,人们对使用无热、无毒和无化学物质的食品保鲜方法兴趣大增。与此同时,市场上也出现了对新鲜、微加工和营养食品的需求。密相二氧化碳(DPCD)是一种在低于标准巴氏杀菌温度下灭活酶和微生物的非热方法,可避免热效应。在过去的几十年里,人们进行了大量研究,以了解其机理及其对细菌、酵母、霉菌和酶等微生物的影响。在许多液态食品中,这项技术都取得了良好的效果。此外,研究人员还在研究 DPCD 与其他非热工艺(如辐照、PEF、超声波和 HPP)的结合效果。本综述汇编了有关 DPCD 的大量研究成果,概述了 DPCD 加工设备、机理、在液态和固态食品中的应用、DPCD 在跨栏技术中的新兴应用,并重点介绍了它对微生物和酶的功效及其在食品加工业中的应用。DPCD 为更好地控制微生物、在更软的工艺设置下灭活酶以及提高质量和营养保留提供了新的机遇。
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引用次数: 0
Review on effect of innovative technologies on shelf-life extension of non-dairy sources from plant matrices 创新技术对延长植物基质非乳制品货架期的影响综述
Pub Date : 2024-07-17 DOI: 10.1016/j.focha.2024.100781

This review covers the nutritional, environmental, and technological aspects of plant-based milk and presents a convincing case for it as a dairy substitute. Almonds, soybeans, oats, coconuts, and other plants are the sources of plant-based milk, which has a variety of tastes, textures, and nutritional profiles. Although plant-based milks can be fortified to match the nutritional profile of dairy milks, dairy milks are known for their high protein, vitamin D, and calcium content. In addition to non-thermal techniques like high-pressure processing, pulsed electric fields, high-intensity ultrasound, and microfluidization, the review explores new thermal technologies like ohmic heating and microwave heating. These technologies are essential for improving the shelf life and inactivating microorganisms and enzymes, which improves the safety and quality of food. In addition, the environmental sustainability of plant-based milk production is examined, emphasizing the possibility of lower resource consumption and greenhouse gas emissions in comparison to dairy milk. Plant-based milk presents a promising substitute for consumers looking for environmentally friendly options or who are subject to dietary restrictions such as lactose intolerance or milk allergies. To maximize the flavor and functionality of plant-based milk products and secure their acceptance in the market, innovative processing techniques are crucial.

这篇综述涵盖了植物乳的营养、环境和技术方面,并提出了植物乳作为乳制品替代品的令人信服的理由。杏仁、大豆、燕麦、椰子和其他植物是植物乳的来源,植物乳有各种口味、质地和营养成分。虽然植物奶可以通过强化达到与牛奶相同的营养成分,但牛奶以蛋白质、维生素 D 和钙含量高而著称。除了高压加工、脉冲电场、高强度超声波和微流控等非热技术外,本综述还探讨了欧姆加热和微波加热等新热技术。这些技术对于延长保质期、灭活微生物和酶,从而提高食品安全和质量至关重要。此外,还对植物奶生产的环境可持续性进行了研究,强调与牛奶相比,植物奶的资源消耗和温室气体排放量更低。植物乳为寻求环保选择的消费者或有乳糖不耐症或牛奶过敏等饮食限制的消费者提供了一种前景广阔的替代品。为了最大限度地提高植物奶产品的风味和功能,确保其被市场接受,创新的加工技术至关重要。
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引用次数: 0
Characterization of bioactive compounds, antioxidants and antimicrobial properties of Allagoptera leucocalyx Allagoptera leucocalyx 的生物活性化合物、抗氧化剂和抗菌特性的表征
Pub Date : 2024-07-13 DOI: 10.1016/j.focha.2024.100775
Ayelen J. Camacho Crespo , Natalia Montellano Duran

Allagoptera leucocalyx is a non-studied tropical fruit distributed in the Bolivian Chiquitania region. This study aimed to investigate its bioactive compounds as well as its antioxidant and antimicrobial properties to encourage its consumption and explore potential applications in various industries. The bioactive compounds: polyphenols, flavonoids, and anthocyanins were determined by the Folin-Ciocalteu, AlCl3, and differential pH methods. Antioxidant activity was assessed by the DPPH and ABTS assays, while antimicrobial activity by turbidimetry and antibiogram. The highest results obtained revealed significant levels of polyphenols (209 ± 12 mg GAE/100 g), and flavonoids (7.9 ± 0.1 mg QE/100 g). Moreover, the fruit exhibit antioxidant activity (9.3 ± 0.2 μmol Trolox/g by ABTS and 5.9 ± 0.2 μmol Trolox/g by DPPH) and antimicrobial activity against Shigella sp. In conclusion, these promising findings suggest that this fruit could be utilized in the development of new products within the cosmetic, food, and pharmaceutical industries.

Allagoptera leucocalyx 是一种未经研究的热带水果,分布在玻利维亚奇基塔尼亚地区。本研究旨在调查其生物活性化合物及其抗氧化和抗菌特性,以鼓励食用并探索其在各行业的潜在应用。生物活性化合物:多酚、类黄酮和花青素采用 Folin-Ciocalteu、AlCl3 和 pH 差值法进行测定。抗氧化活性通过 DPPH● 和 ABTS● 法进行评估,抗菌活性通过浊度法和抗生素图谱法进行评估。结果显示,多酚(209 ± 12 毫克 GAE/100 克)和类黄酮(7.9 ± 0.1 毫克 QE/100 克)的含量最高。此外,这种水果还具有抗氧化活性(ABTS●法为 9.3 ± 0.2 μmol Trolox/g,DPPH●法为 5.9 ± 0.2 μmol Trolox/g)和对志贺氏菌的抗菌活性。
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引用次数: 0
Influence of heat treatment and calcium sequestering salts on in vitro protein digestion in blends of skimmed buffalo and bovine milk 热处理和钙盐对脱脂水牛奶和牛乳混合物体外蛋白质消化的影响
Pub Date : 2024-07-11 DOI: 10.1016/j.focha.2024.100779

This study investigated the impact of the addition of 5 mM trisodium citrate (TSC) or disodium hydrogen phosphate (DSHP) and heat treatment (85 or 95 °C for 5 min) on the in vitro gastrointestinal digestibility of proteins in blends (0:100, 25:75, 50:50, 75:25, 100:0) of skimmed buffalo and bovine milk. Digestibility of proteins decreased with increasing proportion of buffalo skim milk, irrespective of heat treatment and calcium sequestering salt (CSS) addition. Heat treatment alone did not induce significant changes in the digestibility of proteins. CSS addition and heat treatment did not significantly affect whey protein digestibility but impacted the digestibility of κ-casein. While addition of TSC and DSHP induced comparable κ-casein digestibility in bovine skim milk and milk blends, the addition of TSC and heat treatment of buffalo skim milk effected maximum κ-casein gastrointestinal digestibility. Overall, α-lactalbumin and κ-casein were the most resistant proteins against degradation by digestive enzymes. These results may be useful in deciding appropriate process intervention that could maximise gastrointestinal digestion of proteins in buffalo and bovine milk blends.

本研究调查了添加 5 mM 柠檬酸三钠(TSC)或磷酸氢二钠(DSHP)以及热处理(85 或 95 °C 5 分钟)对脱脂水牛奶和牛乳混合物(0:100、25:75、50:50、75:25、100:0)中蛋白质的体外胃肠道消化率的影响。蛋白质的消化率随着脱脂水牛奶比例的增加而降低,与热处理和钙封存盐(CSS)的添加无关。单独进行热处理不会引起蛋白质消化率的显著变化。添加 CSS 和热处理对乳清蛋白的消化率影响不大,但会影响κ-酪蛋白的消化率。在牛脱脂乳和混合乳中添加TSC和DSHP可诱导相似的κ-酪蛋白消化率,而添加TSC和对水牛脱脂乳进行热处理可影响最大的κ-酪蛋白胃肠道消化率。总之,α-乳白蛋白和κ-酪蛋白是最难被消化酶降解的蛋白质。这些结果可能有助于决定适当的加工干预措施,从而最大限度地提高水牛奶和牛乳混合物中蛋白质的胃肠道消化率。
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引用次数: 0
Mushroom-legume-based alternative chicken nuggets: Physico-chemical and sensory properties 蘑菇-豆荚替代鸡块:物理化学和感官特性
Pub Date : 2024-07-09 DOI: 10.1016/j.focha.2024.100777

The consumer demand for meat analogs (MAs) is on the rise. Soy proteins are frequently used as the primary ingredient in MAs, but soybean may negatively impact human health and cognitive functions. The aim of this research was to develop alternative chicken nuggets using mushroom and chickpea flour. Effect of different sources of mushroom and starch in the textural and sensory quality of nuggets were determined. Phoenix mushroom was chosen based on nuggets' textural and sensory qualities. Different ratios of mushroom and chickpea flour (0:70, 15:55, 55:15, and 70:0, w/w) were tested to prepare the nuggets. Based on protein, textural and sensory qualities compared to chicken and commercial plant-based nuggets, the mushroom to flour ratio of 55:15 was chosen. The results showed that tapioca starch-containing nuggets had a better texture and were more palatable to consumers. The processed nuggets demonstrated lower protein (6.88 vs 11.72%, dry wt. basis) but better cooking yield (87.65 vs 85.29%), moisture retention (77.86 vs 75.16%), fat retention (90.55 vs 88.39%) and higher consumer acceptability than commercially available soy-based nuggets. The results of the study suggest that mushroom and chickpea flour may replace soy protein in meat analogs.

消费者对肉类类似物(MAs)的需求在不断增加。大豆蛋白经常被用作类似肉类食品的主要成分,但大豆可能会对人体健康和认知功能产生负面影响。本研究旨在利用蘑菇和鹰嘴豆粉开发替代鸡块。研究确定了不同来源的蘑菇和淀粉对鸡块质地和感官质量的影响。根据鸡块的质地和感官质量选择了凤凰蘑菇。试验了不同比例的蘑菇和鹰嘴豆粉(0:70、15:55、55:15 和 70:0,重量比)来制备蘑菇块。根据蛋白质、质地和感官质量与鸡肉块和商业植物块的比较,选择了 55:15 的蘑菇和面粉比例。结果表明,含木薯淀粉的鸡块口感更好,消费者也更喜欢。加工后的蘑菇块蛋白质含量较低(6.88% 对 11.72%,干重),但烹饪率(87.65% 对 85.29%)、水分保持率(77.86% 对 75.16%)、脂肪保持率(90.55% 对 88.39%)和消费者接受度均高于市售大豆蘑菇块。研究结果表明,蘑菇和鹰嘴豆粉可以替代大豆蛋白制作肉类类似物。
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引用次数: 0
CRISPR/Cas9-mediated gene editing of L. bulgaricus d-lactate dehydrogenase (ldh) gene to monitor yogurt characteristics CRISPR/Cas9 介导的保加利亚乳杆菌 d-乳酸脱氢酶(ldh)基因编辑用于监测酸奶特性
Pub Date : 2024-07-08 DOI: 10.1016/j.focha.2024.100772
G. M. M. Anwarul Hasan , Anuj Kumer Das , Mohammed A. Satter

Yogurt is a coagulated dairy product that results from the fermentation of milk and milk products by lactic acid bacteria, specifically Lactobacillus bulgaricus and Streptococcus thermophilus. Clustered regularly interspaced short palindromic repeats (CRISPR)/Cas9 is an emerging tool in molecular biology research which can be used extensively in prokaryotes and eukaryotes. Lactate dehydrogenase (LDH) is an important enzyme of glycolytic pathway. In this research, we have edited L. bulgaricus d-lactate dehydrogenase (ldh) gene using CRISPR/Cas9 system. Genome editing was accomplished through synthesizing gDNA on the basis of PAM region of ldh gene and ligation of synthesized gDNA with pCRISPR-SacB plasmid which was later co-transformed with mutagenic donor DNA into competent cells containing pCasRed. This method combined CRISPR/Cas9 with γ Red machinery to achieve editing of ldh gene. The PCR screening and sequencing results confirmed that, base deletion has been occurred in the target region of ldh gene. Differences in several characteristics including texture, pH were observed on the yogurt samples from newly developed L. bulgaricus strains compared to normally produced yogurt. Our results suggest that, other genes of lactobacillus can be edited through CRISPR/Cas9 technology which can be extensively used for metabolic engineering.

酸奶是由乳酸菌(特别是保加利亚乳杆菌和嗜热链球菌)对牛奶和奶制品进行发酵后凝固而成的乳制品。聚类规则间隔短回文重复序列(CRISPR)/Cas9 是分子生物学研究中的一种新兴工具,可广泛用于原核生物和真核生物。乳酸脱氢酶(LDH)是糖酵解途径中的一种重要酶。在这项研究中,我们利用 CRISPR/Cas9 系统编辑了保加利亚乳杆菌的 d-乳酸脱氢酶(ldh)基因。基因组编辑是以ldh基因的PAM区为基础合成gDNA,然后将合成的gDNA与pCRISPR-SacB质粒连接,再将质粒与诱变供体DNA共同转入含有pCasRed的合格细胞中。这种方法结合了 CRISPR/Cas9 和 γ Red 机制,实现了对 ldh 基因的编辑。PCR筛选和测序结果证实,ldh基因的靶区发生了碱基缺失。与正常生产的酸奶相比,新培育的保加利亚乳杆菌菌株生产的酸奶样品在质地、pH 值等方面存在差异。我们的研究结果表明,可以通过 CRISPR/Cas9 技术编辑乳酸菌的其他基因,该技术可广泛用于代谢工程。
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Food chemistry advances
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