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旨在高值化利用青刺果榨油后副产物青刺果粕中的蛋白质，以青刺果粕为原料，提取青刺果蛋白并制备蛋白酶解物，采用活菌吸附法联合反相高效液相色谱（RP-HPLC）从蛋白酶解物中筛选出抗菌肽组分，评价其抑菌活性，并对抑菌活性最好的组分进行稳定性分析。结果表明：筛选出的3个抗菌肽组分（F1、F2和F3）中，F2组分抑菌活性最好，当其质量浓度为10 mg/mL时对金黄色葡萄球菌和大肠杆菌的抑菌活性分别达到45.5%和39.1%；抗菌肽组分F2抑菌活性随温度升高和盐溶液浓度增加均降低，最适pH为6.0，在酸性环境中极不稳定，对酸性蛋白酶、碱性蛋白酶、胃蛋白酶和胰蛋白酶的耐受性均较差，对中性蛋白酶耐受性较好。综上，青刺果粕中的蛋白酶解物具有抑菌活性，具有开发青刺果抗菌肽的潜力。In order to make high value use of the proteins in the by-products of Prinsepia utilis Royle oil extraction, Prinsepia utilis Royle proteins were extracted from the meal of Prinsepia utilis Royle and then hydrolyzed to prepare hydrolysates. The antimicrobial peptide fractions were screened from the hydrolysates by using live bacterial adsorption combined with reversed-phase high-performance liquid chromatography (RP-HPLC), their antibacterial activities were evaluated, and the stability of the fraction with the best antibacterial activity was analyzed. The results showed that among the three antimicrobial peptide fractions (F1, F2 and F3) screened, the F2 fraction had the best antibacterial activity, and its antibacterial activities against Staphylococcus aureus and Escherichia coli reached 45.5% and 39.1%, respectively, at a mass concentration of 10 mg/mL. The antibacterial activity of the antimicrobial peptide fraction F2 was reduced with the increase of temperature and salt solution concentration, and its optimal pH was 6.0, and it was extremely unstable in an acidic environment. The antimicrobial peptide fraction F2 was poorly tolerated by acid protease, alkaline protease, pepsin and trypsin, and better tolerated by neutral protease. In conclusion, the enzymatic hydrolysates of protein in the meal of Prinsepia utilis Royle have antibacterial activity, and has potential for the development of antimicrobial peptides from Prinsepia utilis Royle.

旨在促进辣木籽醒酒肽的开发利用，以辣木籽蛋白粉为原料，蛋白水解度和辣木籽醒酒肽的乙醇脱氢酶(ADH)激活率为考察指标，采用单因素实验和响应面法优化辣木籽醒酒肽的制备工艺。结果表明，辣木籽醒酒肽制备的最佳酶解工艺条件为料液比1∶ 20、碱性蛋白酶添加量5.0%、酶解pH 9.0、酶解温度63 ℃、酶解时间142 min，在此条件下辣木籽蛋白水解度为12.23%，辣木籽醒酒肽的ADH激活率为22.16%。综上，以辣木籽为原料制备醒酒肽具有良好的发展前景。To promote the development and utilization of Moringa oleifera seed sobering peptide, Moringa oleifera seed protein powder was used as raw material, and the hydrolysis degree of protein and the alcohol dehydrogenase (ADH) activation rate of sobering peptide were used as indicators, single factor experiment and response surface methodology were used to optimize the preparation process of Moringa oleifera seed sobering peptide. The results showed that the optimum enzymatic hydrolysis conditions for the preparation of Moringa oleifera seed sobering peptide were as follows: solid-liquid ratio 1∶ 20, addition amount of alkaline protease 5.0%, enzymatic hydrolysis pH 9.0, enzymatic hydrolysis temperature 63 ℃, and enzymatic hydrolysis time 142 min. Under these conditions, the hydrolysis degree of Moringa oleifera seed protein was 12.23%, and the ADH activation rate of Moringa oleifera seed sobering peptide was 22.16%. In conclusion, the preparation of sobering peptide from Moringa oleifera seed has a good prospect for development.

为给辣木籽凝乳酶的开发利用提供理论依据，以脱脂辣木籽粉为原料，采用盐法提取、硫酸铵（AS）分级沉淀和离子交换色谱法提取分离凝乳酶。通过单因素试验和正交试验对辣木籽凝乳酶的盐法提取条件进行优化，采用单因素试验对离子交换色谱法分离纯化条件进行优化，并采用SDS-PAGE和RP-HPLC对分离得到的辣木籽凝乳酶的分子质量和纯度进行分析。结果表明：盐法提取的最佳条件为料液比1∶ 10、提取温度30 ℃、pH 7.15、NaCl浓度0.3 mol/L、提取时间0.5 h；在20%~40% AS饱和度下的沉淀蛋白凝乳活性最好且蛋白质总占有率达6171%；离子交换色谱法的最佳分离条件为进样质量浓度30 mg/mL、流动相pH 5.15、流速1.68 mL/min、流动相组成为0.05 mol/L 醋酸钠+0.05 mol/L氯化钠，在此条件下辣木籽凝乳酶的凝乳比活力值达到24 SU/mg；该凝乳酶的分子质量主要分布在35～45 kDa，纯度达到90%以上。采用该方法能有效提取分离辣木籽凝乳酶，且能很好地保持其凝乳活性。 To provide theoretical basis for the development and utilization of milk-clotting protease from Moringa oleifera seeds, the milk-clotting protease were extracted from defatted Moringa oleifera seed powder by salt method, and separated by ammonium sulfate (AS) fractional precipitation and ion exchange chromatography. The salt extraction conditions of milk-clotting protease from Moringa oleifera seeds were determined by single factor experiment and orthogonal experiment, and the ion exchange chromatography separation conditions were optimized by single factor experiment. The molecular weight and purity of the milk-clotting protease from Moringa oleifera seeds were analyzed using SDS-PAGE and RP-HPLC. The results showed that the optimal extraction conditions of salt method were obtained as follows: solid-liquid ratio 1∶ 10, extraction temperature 30 ℃, pH 7.15, sodium chloride concentration 0.3 mol/L and extraction time 0.5 h. Under 20%-40% AS saturation, the precipitated protein had the best milk coagulating activity and the total percentage of extracted protein reached 61.71%. The optimal separation conditions of ion exchange chromatography were obtained as follows: injection mass concentration 30 mg/mL, mobile phase pH 5.15, flow rate 1.68 mL/min, and mobile phase consisted of 0.05 mol/L sodium acetate and 0.05 mol/L sodium chloride. Under these conditions, the milk coagulating specific activity value of the milk-clotting from Moringa oleifera seeds reached 24 SU/mg.The molecular weight of milk-clotting protease was mainly distributed in the range of 35-45 kDa, and the purity was over 90%. This method can effectively extract and separate milk-clotting protease from Moringa oleifera seeds, and maintain its milk coagulating activity well.

旨在为辣木籽抗菌肽的储存、加工提供参考，以实验室前期筛选的具有较好抑制金黄色葡萄球菌生长的辣木籽肽F2和F3为原料，测定其在不同储存、加工条件下的相对抑菌率，分析抑菌稳定性。结果表明：低温储存有利于辣木籽抗菌肽F2和F3抑菌活性的保留，且储存温度为-20 ℃时，其抑菌活性受储存时间影响较小；F2、F3随pH增大相对抑菌率先升高后降低，F2、F3适宜的pH分别为6和8；加工温度、超声波功率升高，F2、F3相对抑菌率降低；相比于Na+和K+，Mg2+对F2、F3相对抑菌率的降低作用更大。实际加工和储存过程中应选择合适的条件，以保证辣木籽抗菌肽抑菌活性的保留。 In order to provide reference for the storage and processing of antimicrobial peptides from Moringa oleifera seeds, the Moringa oleifera seed peptide F2 and F3 with good anti-Staphylococcus aureus activity screened in the laboratory were used as raw materials to determine their relative bacterial inhibition rates under different storage and processing conditions and to analyze antimicrobial stability. The results showed that low temperature storage was beneficial to the rentention of antimicrobial acitivity of F2,F3, and the antimicrobial activity was less affected by the storage time when the storage temperature was -20 ℃. The relative bacterial inhibition rate of F2 and F3 increased and then decreased with the increase of pH, and the suitable pH of F2 and F3 were 6 and 8, respectively. The relative bacterial inhibition rate of F2 and F3 decreased with the increase of processing temperature and ultrasonic power. Compared with Na+ and K+, Mg2+ had a greater effect on reducing the relative bacterial inhibition rate of F2 and F3. The suitable conditions should be selected during actual processing and storage to ensure the retention of the antimicrobial activity of Moringa oleifera seeds antimicrobial peptide.

为促进辣木籽油中神经酸的开发利用，以辣木籽为原料，采用低温压榨法、超临界CO2萃取法、索氏抽提法提取辣木籽油，筛选合适的提油方法，再以辣木籽油为原料制备混合脂肪酸，通过低温结晶法纯化辣木籽油中的神经酸，通过单因素实验和响应面实验优化神经酸的纯化工艺。结果表明：低温压榨法提取的辣木籽原油中神经酸含量较高，达到0.88%；低温结晶法纯化辣木籽油中神经酸的最优工艺条件为料液比（混合脂肪酸与结晶溶剂质量体积比）1∶ 4.2、结晶温度-21 ℃、结晶时间2.3 h，该条件下所得产品中神经酸含量为(5.36±0.01)%，相比辣木籽原油中神经酸含量提升了5.1倍。通过低温压榨法提取辣木籽油，再采用低温结晶法纯化神经酸，能够获得神经酸含量较高的产品。 In order to promote the development and utilization of nervonic acid in Moringa oleifera seed oil, with Moringa oleifera seed as raw material, Moringa oleifera seed oil was extracted by low temperature pressing, supercritical CO2 extraction and Soxhlet extraction to select suitable oil extraction method. Then, mixed fatty acids were prepared using Moringa oleifera seed oil as raw material, and the nervonic acid in Moringa oleifera seed oil was purified by low temperature crystallization method. The purification process of nervonic acid was optimized by single factor experiment combined with response surface methodology. The results showed that the content of nervonic acid in crude Moringa oleifera seed oil extracted by low temperature pressing method was higher, reaching 0.88%.The optimal purification process of nervonic acid from Moringa oleifera seed oil by low temperature crystallization method was obtained as follows: solid-liquid ratio(ratio of mixed fatty acid mass to crystallization solvent volume) 1∶ 4.2, crystallization temperature -21 ℃ and crystallization time 2.3 h. Under these conditions, the nervonic acid content in the product was (5.36±0.01)%, which was 5.1 times higher than that in crude Moringa oleifera seed oil. By extracting Moringa oleifera seed oil through low temperature pressing method and purifying nervonic acid through low temperature crystallization method, products with high levels of nervonic acid can be obtained.

为进一步提高辣木籽蛋白资源的开发利用，采用盐提法提取辣木籽蛋白，再采用超声波辅助酶法制备辣木籽ACE抑制肽。以水解度和ACE抑制率为评价指标，通过单因素实验探究超声波功率、超声酶解时间、超声酶解温度及料液比对制备ACE抑制肽的影响,采用响应面法对制备工艺条件进行优化。结果表明：超声波辅助酶法制备辣木籽ACE抑制肽的最佳酶解工艺条件为碱性蛋白酶添加量5.5 mg/mL、pH 9、超声波功率500 W、超声酶解时间1.7 h、超声酶解温度55 ℃、料液比1∶ 45，在此条件下制备的酶解物ACE抑制率达到78.32%，水解度为778%。以辣木籽为原料制备ACE抑制肽作为功能性蛋白肽产品，可有效提高辣木籽蛋白资源的开发利用。 To further improve the exploitation of Moringa oleifera seed protein resources, the protein in Moringa oleifera seeds was extracted by salt extraction method, and then Moringa oleifera seeds ACE inhibitory peptide was prepared by ultrasound-assisted enzymatic hydrolysis. With the degree of hydrolysis and ACE inhibition rate as the evaluation indexes, the effects of the ultrasonic power, ultrasonic enzymolysis time, ultrasonic enzymolysis temperature and ratio of material to liquid on the preparation of ACE inhibitory peptide were studied through single factor experiment. The preparation process conditions of ACE inhibitory peptide were optimized by response surface methodology. The results showed that the optimal ultrasound-assisted enzymatic hydrolysis conditions were obtained as follows: Alkaline protease dosage 5.5 mg/mL,pH 9,ultrasonic power 500 W, ultrasonic enzymolysis time 1.7 h, ultrasonic enzymolysis temperature 55 ℃ and ratio of material to liquid 1∶ 45. Under these conditions, the ACE inhibition rate of the prepared product reached 78.32% and the degree of hydrolysis was 7.78%. Using ACE inhibitory peptide prepared from Moringa oleifera seeds as a functional protein peptide product can effectively improve the exploitation of Moringa oleifera seed protein resources.

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制备大肠杆菌细胞膜固定相(E.coli cell membrane stationary phase，ECMSP)，研究其对辣木籽抗菌肽的萃取效果。研究超声时间、超声功率对大肠杆菌破壁效果的影响，分析大肠杆菌细胞膜与大孔硅胶的吸附特征及ECMSP对辣木籽抗菌肽的吸附效果，明确萃取前后抑菌活力变化及反相-高效液相色谱（RP-HPLC）差异峰。结果表明：在超声功率500 W、超声时间30 min条件下，大肠杆菌破壁效果较好；大孔硅胶对大肠杆菌细胞膜的饱和吸附值（Csmax）为9.98 μg/mg、吸附常数（K*）为171.02 μg/mL；萃取前后抗菌肽对大肠杆菌和金黄色葡萄球菌的抑制效果差异明显，RP-HPLC分析表明萃取前后出现了3个差异峰。研究结果可为辣木籽抗菌肽的后续开发利用提供理论基础和应用参考。 E. coli cell membrane stationary phase (ECMSP) was prepared, and its extraction effect of Moringa oleifera seed antimicrobial peptides was studied. The effects of ultrasonic time and ultrasonic power on the wall breaking effect of E. coli were studied. The adsorption characteristics of E. coli cell membrane and macroporous silica gel, and the adsorption effect of ECMSP on the Moringa oleifera seed antimicrobial peptides were analyzed. The change of antibacterial activity and the reverse-high performance liquid chromatography (RP-HPLC) difference peaks before and after extraction were clarified. The results showed that when the ultrasonic power was 500 W and the ultrasonic time was 30 min, the wall breaking effect was good. The saturated adsorption value （Csmax）of the macroporous silica gel to the cell membrane of E. coli was 9.98 μg/mg, and the adsorption constant (K*) was 171.02 μg/mL. The inhibitory effects of antimicrobial peptides on Escherichia coli and Staphylococcus aureus were significantly different, and RP-HPLC analysis showed three different peaks before and after extraction. The research could provide theoretical basis and application references for the subsequent development and utilization of Moringa oleifera seed antimicrobial peptides.

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目的总结2021年11月1日—2022年10月31日沈阳地区呼吸道病毒感染的流行病学特征。方法 2021年11月1日—2022年10月31日沈阳市儿童医院收治的急性呼吸道感染患儿3206例,取患儿咽拭子标本采用 多重RT-PCR法检测常见的10种呼吸道病毒,取患儿外周血采用ELISA法检测血清EB病毒(EBV)抗体。结果 3206 例患儿中至少1种呼吸道病毒阳性率55.65%,病原体分布情况依次为乙型流感病毒(Infb)感染18.53%, 合胞病毒 (RSV)感染12.45%, 鼻病毒(HRV)感染9.01%, 甲型流感病毒(Infa)感染6.14%, 偏肺病毒(HMPV)感染5.68%, EBV感染3.59%, 副流感病毒(HIV)感染2.40%, 腺病毒(HADV)感染1.59%, 冠状病毒(HCOV)感染0.90%, 博卡 病毒(BoV)感染0.47%。与其他季节比较,冬季Infb感染阳性率高,春冬季RSV感染阳性率高,夏季HRV､Infa及 HMPV感染阳性率高(P均<0.05)。不同性别, 年龄ARTI患儿呼吸道病毒感染阳性率比较差异均无统计学差异(P 均>0.05)。结论 2021年11月1日—2022年10月31日间沈阳地区儿童急性呼吸道感染主要以Infb, RSV, HRV, Infa､HMPV感染为主。儿童急性呼吸道病毒感染与季节有关,Infb感染主要集中在冬季,RSV感染主要集中在冬春 季,HRV感染主要集中在夏秋季,Infa感染主要集中在夏季。 [ABSTRACT FROM AUTHOR]

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