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Enzymatic hydrolysis as a valorization strategy of bovine lungs: Optimization of process variables and study of antioxidant capacity 酶水解作为牛肺的增值策略:过程变量的优化和抗氧化能力的研究
Pub Date : 2023-03-21 DOI: 10.1002/jsf2.110
F. G. Martinez, V. A. Ambrosi, G. Rocha, A. M. Sancho, N. Szerman

Background

The increase in meat production would also imply an increase in the generation of by-products or edible offal. These by-products can be used, due to their high protein content, as a substrate to obtain hydrolysates with functional activities. This study aimed to valorize bovine lungs by producing hydrolysates with antioxidant capacity.

Results

A response surface methodology study was carried out using a central composite design in which factors were temperature (43.2–76.8°C), enzyme/substrate ratio [ES: 0.65–4.85% (w/w)] using Alcalase 2.4 L/Flavourzyme (1:1) and pH (5.8–9.2). The antioxidant capacity of the obtained BLH (bovine lung hydrolysates) was evaluated by the ABTS, DPPH, and FRAP methods. The maximum antioxidant capacity was achieved at 53.°C, ES ratio of 2.3% (w/w), and pH of 8.2. Under optimal hydrolysis conditions, the antioxidant capacity increased rapidly from the beginning of the reaction to 30 min, reaching a maximum at 120 min; then, it decreased until the end (180 min). The degree of hydrolysis increased as hydrolysis occurred up to a maximum of 45%. Molecular weight distribution of BLH evaluated by Tricine-SDS-PAGE and SEC-FPLC showed that bovine lung proteins hydrolyzed for 120 min presented low molecular weight peptides (<5 kDa), which are absent in the non-hydrolyzed sample.

Conclusion

In this work, we optimized the variables for producing BLH with antioxidant capacity using commercial proteases in a short reaction time, making it a worthwhile strategy for the recovery of by-products from the meat industry.

肉类产量的增加也意味着副产品或可食用内脏的增加。由于蛋白质含量高,这些副产物可以作为底物来获得具有功能活性的水解产物。本研究旨在通过生产具有抗氧化能力的水解物来激活牛肺。结果采用中心复合设计,采用响应面法研究温度(43.2 ~ 76.8℃)、酶/底物比(ES: 0.65 ~ 4.85% (w/w)) (Alcalase∶2.4 L/Flavourzyme∶1:1)和pH(5.8 ~ 9.2)。通过ABTS、DPPH和FRAP方法评价所得BLH(牛肺水解物)的抗氧化能力。在53岁时达到最大抗氧化能力。°C, ES比2.3% (w/w), pH为8.2。在最佳水解条件下,从反应开始到反应30 min,抗氧化能力迅速增加,在反应120 min时达到最大值;然后逐渐减小,直至结束(180 min)。水解的程度随着水解的发生而增加,最高可达45%。通过Tricine-SDS-PAGE和SEC-FPLC对BLH的分子量分布进行了分析,结果表明,水解120 min的牛肺蛋白中存在低分子量肽(< 5kda),而非水解样品中不存在。结论本研究优化了利用商业蛋白酶在短时间内生产具有抗氧化能力的BLH的工艺参数,为肉类工业副产品的回收提供了一个有价值的策略。
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引用次数: 0
Bioactive compounds, functional properties, and technological application of Passiflora quadrangularis: A review 西番莲的生物活性成分、功能特性及技术应用综述
Pub Date : 2023-03-15 DOI: 10.1002/jsf2.108
Josias M. A. Cruz, Zilanir C. Pereira, Renilto F. Corrêa, Carlos V. Lamarão, Edgar A. Sanches, Pedro H. Campelo, Jaqueline de A. Bezerra

The objective of this research was to evaluate the bioactive compounds and functional properties of Passiflora quadrangularis, aiming to contribute to future studies on the elaboration of functional products based on this species. The chemical analysis of flavonoids from the leaves, as well as the chemical composition of fruits (specifically the pericarp) were accessed herein. The functional properties are directly dependent on specific extracts, and the aqueous extract and the flavonoid fraction are responsible for the sedative property. The presence of C-glycosyl flavonoids is highly reported in the chemical composition, highlighting the vitexin-2″-O-xyloside. Few studies have been focused on this species, and most of them are devoted to the sedative properties of the genus Passiflora spp. However, a review on the main available information on this species is of extreme interest for the research and development of products in pharmaceuticals, cosmetics, and food areas.

本研究的目的是对西番莲的生物活性成分和功能特性进行评价,为进一步开发西番莲功能性产品提供依据。本文分析了叶中黄酮类化合物的化学成分,以及果实(特别是果皮)的化学成分。其功能特性直接依赖于特定的提取物,其中水提取物和黄酮类化合物部分具有镇静作用。c -糖基类黄酮的存在在化学成分中被高度报道,突出的是牡荆素-2″- o -木糖苷。目前对西番莲属植物的研究较少,主要集中在西番莲属植物的镇静作用方面,但对西番莲属植物的主要研究资料进行综述,对西番莲属植物在医药、化妆品和食品等领域的产品研究和开发具有重要意义。
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引用次数: 0
Enzyme-assisted HPTLC method for the simultaneous analysis of inositol phosphates and phosphate 酶辅助HPTLC法同时分析肌醇磷酸和磷酸
Pub Date : 2023-03-15 DOI: 10.1002/jsf2.109
Corinna Henninger, Bernd Spangenberg, Mario Schmidt, Katrin Ochsenreither, Thomas Eisele

Background

The analysis of myo-inositol phosphates (InsPx) released by phytases during phytic acid degradation is challenging and time-consuming, particularly in terms of sample preparation, isomer separation, and detection. However, a fast and robust analysis method is crucial when screening for phytases during protein engineering approaches, which result in a large number of samples, to ensure reliable identification of promising novel enzymes or target variants with improved characteristics, for example, pH range, thermal stability, and phosphate release kinetics.

Results

The simultaneous analysis of several InsPx (InsP1-InsP4 and InsP5 + 6) as well as free phosphate was established on cellulose HPTLC plates using a buffered mobile phase. Inositol phosphates were subsequently stained using a novel enzyme-assisted staining procedure. Immobilized InsPx were hydrolyzed by a phytase solution of Quantum® Blueliquid 5G followed by a molybdate reagent derivatization. Resulting blue zones were captured by DAD scan. The method shows good repeatability (intra-day and intra-lab) with maximum deviations of the Rf value of 0.01. The HPTLC method was applied to three commercially available phytases at two pH levels relevant to the gastrointestinal tract of poultry (pH 5.5 and pH 3.6) to observe their phytate degradation pattern and thus visualize their InsPx fingerprint.

Conclusion

This HPTLC method presents a semi-high-throughput analysis for the simultaneous analysis of phytic acid and the resulting lower inositol phosphates after its enzymatic hydrolysis and is also an effective tool to visualize the InsPx fingerprints and possible accumulations of inositol phosphates.

植酸降解过程中由植酸酶释放的肌醇磷酸(InsPx)的分析是具有挑战性和耗时的,特别是在样品制备、异构体分离和检测方面。然而,在蛋白质工程方法中筛选植酸酶时,快速和强大的分析方法至关重要,这将导致大量的样品,以确保可靠地鉴定有前途的新酶或具有改进特性的靶变异体,例如pH范围,热稳定性和磷酸盐释放动力学。结果采用缓冲流动相,在纤维素HPTLC板上建立了多种InsPx (InsP1-InsP4和InsP5 + 6)和游离磷酸盐的同时分析方法。肌醇磷酸随后用一种新的酶辅助染色方法染色。固定化的InsPx用Quantum®Blueliquid 5G植酸酶溶液水解,然后用钼酸盐试剂衍生化。由此产生的蓝色区域被DAD扫描捕获。该方法重复性好(日间和实验室内),Rf值的最大偏差为0.01。采用HPTLC方法对3种市售植酸酶在与家禽胃肠道相关的两种pH水平(pH 5.5和pH 3.6)下的植酸酶进行分析,观察其植酸降解模式,并绘制其InsPx指纹图谱。结论该方法为同时分析植酸及其酶解后所得的低磷酸肌醇提供了半高通量分析方法,同时也是可视化insx指纹图谱和可能积累的磷酸肌醇的有效工具。
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引用次数: 1
Nutritional value and health benefits of cashew apple 腰果苹果的营养价值和健康益处
Pub Date : 2023-02-17 DOI: 10.1002/jsf2.107
Yaw Gyau Akyereko, Georgina Benewaa Yeboah, Faustina Dufie Wireko-Manu, Francis Alemawor, F. C. Mills-Robertson, William Odoom

Cashew (Anacardium occidentale) has received both local and international recognition due to its expanding production capacity, trading, and nut utilization. The cashew apple, which forms about 90% of the total fruit weight, is left on the farmers' fields as agricultural waste owing to limited knowledge of its health benefits, a lack of technical know-how and processing equipment, and its high perishability and astringent taste. Numerous studies have been conducted exploring the nutrients and contribution of the apple to human nutrition. This paper reviewed recent articles on the nutritional composition and health benefits of cashew apples for easy accessibility and readership. Cashew apple contains good amounts of vitamin C, sugars (fructose and sucrose), fibers, flavonoids, carotenoids, total polyphenols, volatile components, flavanols, amino acids, and minerals, such as potassium, magnesium, sodium, and iron, which are good for maintaining strong immunity, scavenging free radicals, neuropathic functioning, cardiac functioning, and maintenance of body and skin integrity. Cashew apple is associated with weight loss, and is good for diabetic patients due to its high content of flavonoids (myricetin and quercetin). Consumption of cashew apple and its value-added products confer good health, therapeutic effects in the management of diabetics and cardiovascular diseases, and also ensure food and nutrition security. It is hoped that this review will contribute significantly to the expanding body of knowledge aimed at promoting cashew apple utilization globally.

腰果(Anacardium occidentale)因其不断扩大的生产能力、贸易和坚果利用而获得了当地和国际的认可。腰果苹果约占水果总重量的90%,但由于对其健康益处的了解有限,缺乏技术知识和加工设备,以及其易腐烂和涩味,腰果苹果作为农业废物留在农民的田地里。许多研究都在探索苹果的营养成分和对人体营养的贡献。本文回顾了最近关于腰果苹果的营养成分和健康益处的文章,以方便读者阅读。腰果苹果含有大量的维生素C、糖(果糖和蔗糖)、纤维、类黄酮、类胡萝卜素、总多酚、挥发性成分、黄烷醇、氨基酸和矿物质(如钾、镁、钠和铁),对保持强大的免疫力、清除自由基、神经病变功能、心脏功能以及维持身体和皮肤的完整性都有好处。腰果苹果与减肥有关,由于其高含量的黄酮类化合物(杨梅素和槲皮素)对糖尿病患者有好处。食用腰果苹果及其增值产品对健康有益,对糖尿病和心血管疾病有治疗作用,并确保食品和营养安全。希望本文的综述能对扩大腰果在全球的利用知识体系有所贡献。
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引用次数: 0
Physicochemical properties and antioxidant activity of polysaccharides from Chinese yam (Dioscorea opposita Thunb.) 山药多糖的理化性质及抗氧化活性研究
Pub Date : 2023-02-06 DOI: 10.1002/jsf2.105
Ruize Jiao, Yingyin Wu, Fangcheng Xu, Yan Li, Linchun Mao

Background

Nuoshanyao (NSY) is a special species of Chinese yam (Dioscorea opposita Thunb.) and shows rich functionality and superior nutritional value. The chemical components and structural properties of Nuoshanyao polysaccharide (NSYP) were investigated in his study.

Results

The molecular weight (Mw) of the major component in the polysaccharide was 23 kDa. NSYP was mainly composed of glucose, mannose, and galactose, and the molar ratio was 36.5:1.63:1. There might be an ordered helical conformation in NSYP, and the surface of NSYP was granular. NSYP exhibited strong scavenging capacity on the DPPH radical and chelating capacity on the ferrous ion in vitro with a rate of 94.94% and 82.49% at 4 mg/mL.

Conclusion

NSYP with the appropriate molecular weight range and helix structure could possess great antioxidant activity, while monosaccharide compositions may affect the activity. This work supplied important information for the potential application of NSYP in the nutritional and food industries.

糯山药(NSY)是山药(Dioscorea opposita Thunb.)的一种特殊品种,具有丰富的功能性和优越的营养价值。对糯山药多糖的化学成分和结构性质进行了研究。结果多糖中主要成分分子量为23 kDa。NSYP主要由葡萄糖、甘露糖和半乳糖组成,摩尔比为36.5:1.63:1。NSYP可能存在有序的螺旋构象,NSYP表面呈颗粒状。NSYP对DPPH自由基具有较强的清除能力,对亚铁离子具有较强的螯合能力,在4 mg/mL浓度下,其螯合率分别为94.94%和82.49%。结论适当分子量范围和螺旋结构的NSYP具有较强的抗氧化活性,而单糖成分可能影响其抗氧化活性。这项工作为NSYP在营养和食品工业中的潜在应用提供了重要的信息。
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引用次数: 0
Foliar humic acid and salicylic acid application stimulates physiological responses and antioxidant systems to improve maize yield under water limitations 叶片施用腐植酸和水杨酸刺激生理反应和抗氧化系统提高水分限制下玉米产量
Pub Date : 2023-02-06 DOI: 10.1002/jsf2.106
Adnan Altaf, Fahim Nawaz, Sadia Majeed, Muhammad Ahsan, Khawaja Shafique Ahmad, Gulzar Akhtar, Muhammad Asif Shehzad, Hafiz Muhammad Rashad Javeed, Muhammad Farman

Background

Humic acid (HA) is an organic acid that is naturally present in soil organic matter and improves nutrient availability and the mechanisms involved in plant growth and development. Likewise, salicylic acid (SA) is an important plant hormone involved in the regulation of plant growth and development. A pot experiment was carried out to determine the effects of individual or combined HA and SA application on growth and yield of maize (Zea mays L.) under drought stress conditions. Two maize hybrids, namely, 30T60 (drought tolerant) and 75S75 (drought sensitive), were grown in semi-controlled conditions and foliar applied with SA (1 mM), HA (100 mg L−1) and their combination (HA + SA). The plants were exposed to drought stress at the tasseling stage (R1, 60 days after sowing) for 2 weeks, while control plants were given normal irrigation.

Results

The results showed that HA and SA applications significantly enhanced the gas exchange characteristics (photosynthetic rate, transpiration rate, and stomatal conductance), and antioxidant activity (catalase, guaiacol peroxidase, and superoxide dismutase) of water stressed maize plants. Foliar SA spray significantly increased the photosynthetic efficiency and activity of enzymatic antioxidants closely followed by HA + SA application that ultimately improved the yield and net benefit cost ratio of maize under water deficit conditions.

Conclusion

Our findings suggest that foliar spraying of SA at the initiation of the reproductive stage is a cost-effective strategy to obtain a high maize yield under limited water conditions.

腐植酸(HA)是一种天然存在于土壤有机质中的有机酸,能提高养分有效性,并参与植物生长发育机制。同样,水杨酸(salicylic acid, SA)也是一种重要的植物激素,参与调节植物的生长发育。通过盆栽试验,研究了干旱胁迫条件下单独或组合施用HA和SA对玉米生长和产量的影响。在半控制条件下,选用耐旱玉米30T60和干旱敏感玉米75S75,叶面分别施用SA (1 mM)、HA (100 mg L−1)及其组合(HA + SA)。在抽雄期(R1,播种后60天),植株经受干旱胁迫2周,对照植株正常灌溉。结果表明,施用HA和SA显著提高了水分胁迫玉米植株的气体交换特性(光合速率、蒸腾速率和气孔导度)和抗氧化活性(过氧化氢酶、愈创木酚过氧化物酶和超氧化物歧化酶)。叶片喷施SA显著提高了玉米的光合效率和酶促抗氧化剂活性,其次是HA + SA,最终提高了水分亏缺条件下玉米的产量和净效益成本比。结论在有限水分条件下,在玉米繁殖初期叶面喷施SA是提高玉米产量的有效措施。
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引用次数: 3
Formulation of nutritious gluten-free cookies with upcycled pecan nut cake flour 配方营养无麸质饼干与升级再造的山核桃蛋糕粉
Pub Date : 2023-02-05 DOI: 10.1002/jsf2.104
Letícia Frasson Martendal, Maria Luiza Tonetto Silva, Raquel da Silva Simão, Jane Mara Block, Jaqueline Oliveira de Moraes, Ana Paula Gines Geraldo, Maria Manuela Camino Feltes

Background

The pecan nut [Carya illinoinensis (Wangenh.) C. Koch] press cake (PNC) is a co-product of pressing to produce oil, that has low cost and is nutritionally rich, but still little explored. This study evaluated the effects of different concentrations of PNC flour (PNCF) (0%, 15%, and 30%) to obtain nutritious gluten-free cookies based on rice flour and corn starch, and upcycling the high-quality pecan press cake.

Results

The addition of the PNCF significantly increased the cookies lipids, insoluble fibers, and protein contents, and led to a significant reduction of the carbohydrates content. Texture profile analysis showed that the addition of PNCF in the samples formulation increased hardness and decreased cohesiveness of dough, showing better behavior for technological processing. After baking, the cookies added with PNCF showed no significant difference in the expansion factor, and required a much higher cutting force than cookies without PNCF (control formulation), due to the higher concentration of insoluble fibers of the former.

Conclusion

The use of up to 30% PNCF allowed formulating gluten-free cookies with a suitable proximate composition and desired texture characteristics. The findings can help the food industries to develop technologically viable formulations to meet the consumers´ demands and nutritional requirements of individuals with gluten restrictions, as well as to upcycle an oleaginous co-product.

山核桃[Carya illinensis (Wangenh.)]压榨饼(PNC)是压榨生产石油的副产品,成本低,营养丰富,但尚未得到充分开发。本研究评估了不同浓度的PNC面粉(PNCF)(0%、15%和30%)对以米粉和玉米淀粉为原料制作营养丰富的无麸质饼干的影响,并对优质山核桃压榨饼进行了升级利用。结果添加PNCF显著提高了饼干的脂质、不溶性纤维和蛋白质含量,显著降低了饼干的碳水化合物含量。织构分析表明,PNCF的加入提高了面团的硬度,降低了面团的粘结性,表现出更好的工艺加工性能。烘焙后,添加PNCF的饼干的膨胀系数没有显著差异,但由于前者的不溶性纤维浓度更高,因此需要的切削力比未添加PNCF的饼干(对照配方)要大得多。结论:使用高达30%的PNCF可以制作出具有合适近似成分和所需质地特征的无谷蛋白饼干。研究结果可以帮助食品工业开发技术上可行的配方,以满足消费者的需求和麸质限制个人的营养需求,并对含油副产品进行升级回收。
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引用次数: 0
Red tilapia by-product hydrolysates: A new nitrogen source for Bifidobacterium lactis HN019 红罗非鱼副产物水解物:乳酸双歧杆菌HN019的新氮源
Pub Date : 2023-01-24 DOI: 10.1002/jsf2.103
Lu Tong, Dongwei Jiang, Shengjun Chen, Huijuan Zhang, Yan Zhang, Hui Hong, Yongkang Luo, Yuqing Tan

Background

Red tilapia by-products possess ample protein and are either discarded or processed into low-value products. The nitrogen source is the most expensive part of the microbial culture, so finding a cheap alternative can better promote the microbial economy. In this study, different combinations of enzymes were used to hydrolyze the by-products of red tilapia, and its effects on the growth of Bifidobacterium lactis HN019 were investigated.

Results

The results showed that hydrolysate hydrolyzed by enzyme combination 4 (alcalase: neutrase: papain: flavorzyme = 1:1:2:1) (EC4) obtained the highest nitrogen recovery (53.63%) and >2000 Da peptide proportion (5.07%). Hydrolysate hydrolyzed by enzyme combination 2 (alcalase: neutrase: papain: flavorzyme = 2:1:1:1) (EC2) has less hydrophobic amino acids and could improve the growth rate in 10 h–14 h in 50% nitrogen source substitution but had worst viable count after 24 h cultivation.

Conclusion

These results indicated that red tilapia by-product hydrolysate was an excellent nitrogen source substitution and suggested that the hydrophobic amino acids in the nitrogen source might be an essential factor affecting the growth of Bifidobacterium lactis HN019. This research enhanced the economic value of red tilapia by-products, minimized the waste of aquatic resources, and provided directions for the utilization of red tilapia by-products.

红罗非鱼副产品含有丰富的蛋白质,被丢弃或加工成低价值产品。氮源是微生物培养中最昂贵的部分,因此寻找一种廉价的替代品可以更好地促进微生物经济。本研究采用不同的酶组合对红罗非鱼副产物进行水解,研究其对乳酸双歧杆菌HN019生长的影响。结果结果表明,酶组合4 (alcalase: neutrase: pap蛋白酶:flavorenzyme = 1:1:2:1) (EC4)水解产物的氮回收率最高(53.63%),2000 Da肽比例最高(5.07%)。酶组合2 (alcalase: neutrase: pap蛋白酶:flavorenzyme = 2:1:1:1)水解产物(EC2)疏水氨基酸较少,在50%氮源替代条件下10 h - 14 h可提高生长速度,但培养24 h后活菌数最差。结论红罗非鱼副产物水解液是一种很好的氮源替代品,氮源中的疏水氨基酸可能是影响乳酸双歧杆菌HN019生长的重要因素。本研究提高了红罗非鱼副产品的经济价值,最大限度地减少了水产资源的浪费,为红罗非鱼副产品的利用提供了方向。
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引用次数: 0
Enhancement of nutritional, antioxidant and potential antidiabetic properties of fermented maize flour by partial substitution with Curculigo pilosa and sensory attributes of its gruel 竹节草部分替代提高发酵玉米粉的营养、抗氧化和潜在抗糖尿病性能及其粥的感官特性
Pub Date : 2023-01-19 DOI: 10.1002/jsf2.102
Kayode O. Karigidi, Emmanuel S. Akintimehin, Foluso O. Adetuyi, Oreoluwa O. Akinola, Charles O. Olaiya

Background

Curculigo pilosa (CP) is an important component of traditional medicine where it is used to treat diabetes mellitus and its associated complications. Despite this, its use as functional ingredient in foods has not been fully explored. Therefore, this study was designed to determine the effects of substituting fermented maize flour with CP on the potential antidiabetic, antioxidant and nutritional properties of fermented maize flour.

Results

Fermented maize flour was substituted with Curculigo pilosa flour at different percentage (2.5%, 5% and 10%) and it effects was evaluated on the potential antidiabetic, antioxidant and nutritional properties of the composite flours. Thereafter, the sensory attributes of its gruel were determined. The results showed that bioactive compounds, antioxidant and inhibitory abilities against α-amylase and α-glucosidase enzymes were significantly (p < 0.05) increased. Also, there was significantly reduction in the estimated glycemic index and in vitro starch digestibility of the composite flours. Protein, fibre, ash and fat contents were significantly (p < 0.05) enhanced while carbohydrate content was decreased in the flour blends. The mineral contents of the fermented maize flour was significantly (p < 0.05) increased upon substitution with CP. The sensory attribute scores of gruel (Ogi) made from different flour blends showed they were accepted although they were significantly (p < 0.05) lower when compared with the fermented maize flour without CP.

Conclusion

The results show that Curculigo pilosa could serve as functional ingredient in preparation of fermented maize flour.

毛茛(Curculigo pilosa, CP)是传统医学中用于治疗糖尿病及其相关并发症的重要成分。尽管如此,它作为功能性成分在食品中的应用还没有得到充分的探索。因此,本试验旨在研究用CP替代发酵玉米粉对发酵玉米粉潜在的抗糖尿病、抗氧化和营养特性的影响。结果以2.5%、5%和10%的黄皮竹粉替代发酵玉米粉,对复合面粉的潜在抗糖尿病、抗氧化和营养性能进行了评价。然后,确定其粥的感官属性。结果表明,活性成分、抗氧化能力和对α-淀粉酶和α-葡萄糖苷酶的抑制能力显著提高(p < 0.05)。此外,复合面粉的估计血糖指数和体外淀粉消化率也显著降低。混合面粉的蛋白质、纤维、灰分和脂肪含量显著提高(p < 0.05),碳水化合物含量显著降低。与未添加粗CP的发酵玉米粉相比,不同发酵玉米粉混合制成的粥(Ogi)的感官属性评分显著(p < 0.05)降低,但均可接受。结论粗CP可作为发酵玉米粉的功能性成分发酵玉米粉的制备。
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引用次数: 0
Variability and chemical composition of extractives content of woody residues of three European orchard's species: Apricot ( Prunus Armeniaca L.), Olive ( Olea Europea L.) and Orange trees ( Citrus Sinensis L.) 杏(Prunus Armeniaca L.)、橄榄(Olea Europea L.)和橙(Citrus Sinensis L.) 3种欧洲果园树种木本残基提取物含量的变异及化学成分
Pub Date : 2023-01-12 DOI: 10.1002/jsf2.99
M. Bruno
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引用次数: 0
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