CN115896209A - A kind of extraction method of feijoa pectin rich in RG-I high ester - Google Patents

A kind of extraction method of feijoa pectin rich in RG-I high ester Download PDF

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CN115896209A
CN115896209A CN202310012365.5A CN202310012365A CN115896209A CN 115896209 A CN115896209 A CN 115896209A CN 202310012365 A CN202310012365 A CN 202310012365A CN 115896209 A CN115896209 A CN 115896209A
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feijoa
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CN115896209B (en
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王丹
王清明
陈其阳
张猛
龙治坚
张瑜
金雪娇
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Southwest University of Science and Technology
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Abstract

本发明公开了一种富含RG‑I高酯的费约果果胶的提取方法,包括以下步骤:将费约果果肉进行预处理成小块并灭活果胶酶后,使用50mM柠檬酸钠盐溶液在微波炉中提取果胶,再进行冷却、加酶酶解、离心,使用乙醇进行提纯后,干燥得到富含RG‑I高酯果胶。本发明公开的提取方法,利用微波辅助酶解能够将费约果果肉中果胶较充分的提取出来,缩短了酶处理时间,大幅提高了提取效率和产品得率;而且本发明制备的果胶为高酯化度果胶,结构更加完整,RG‑Ι含量为41.38%,粘度较高。The invention discloses a method for extracting feijoa pectin rich in RG‑I high esters, comprising the following steps: pretreating feijoa pulp into small pieces and inactivating pectinase, then using 50mM citric acid Pectin is extracted from the sodium salt solution in a microwave oven, then cooled, enzymatically hydrolyzed, centrifuged, purified with ethanol, and dried to obtain high-ester pectin rich in RG‑I. The extraction method disclosed by the invention can fully extract the pectin in the feijoa pulp by using microwave-assisted enzymolysis, shortens the enzyme treatment time, and greatly improves the extraction efficiency and product yield; and the pectin prepared by the invention It is a high degree of esterification pectin with a more complete structure, RG-Ι content of 41.38%, and a higher viscosity.

Description

一种富含RG-I高酯的费约果果胶的提取方法A kind of extraction method of feijoa pectin rich in RG-I high ester

技术领域technical field

本发明属于果胶提取技术领域,具体涉及到一种富含RG-I高酯的费约果果胶的提取方法。The invention belongs to the technical field of pectin extraction, and in particular relates to an extraction method of feijoa pectin rich in RG-I high ester.

背景技术Background technique

费约果又名巴西番石榴,是一种桃金娘科南美稔属的多年生亚热带常绿灌木类植物,属于我国近几年新兴的一类水果。费约果中富含丰富的营养成分和功能活性成分,如蛋白质、维生素、矿物质和多酚等。据报道,费约果果实中果胶含量达到了20%,因此费约果是果胶的资源宝库,而费约果在我国大面积栽种规模待推广,其加工产品待开发,相关的加工产品特别是果胶提取报道及专利甚少。Feijoa, also known as Brazilian guava, is a perennial subtropical evergreen shrub plant belonging to the genus Myrtaceae South America. It belongs to a class of fruits that have emerged in my country in recent years. Feijoa is rich in nutrients and functional active ingredients, such as protein, vitamins, minerals and polyphenols. According to reports, the pectin content of feijoa fruit has reached 20%, so feijoa fruit is a treasure house of pectin resources, and the large-scale planting of feijoa fruit in my country needs to be promoted, and its processed products need to be developed. Related processed products Especially pectin extraction reports and patents are seldom.

果胶是一种广泛存在于双子叶植物细胞壁胞间层的酸性杂多糖。果胶因其良好的乳化性、胶凝性、稳定性和增稠性而广泛用于食品、医药、化妆品等行业中。同时,果胶也是天然的水溶性膳食纤维,能够降低患癌症、糖尿病、肥胖症的风险,调节人体肠道微环境,降低血脂。高酯果胶是指酯化度(甲基酯化的半乳糖醛酸单位与果胶中总半乳糖醛酸单位的比例)≥50%的果胶,由于其形态结构最接近天然果胶分子,具有良好的应用前景。酯化程度会影响果胶的结构、功能性质以及与其他物质形成复合物的能力等,尽可能的保留果胶的完整结构,提高果胶的性能,为更好的应用于食品等行业具有重要意义。Pectin is an acidic heteropolysaccharide widely present in the intercellular layer of the cell wall of dicotyledonous plants. Pectin is widely used in food, medicine, cosmetics and other industries because of its good emulsification, gelling, stability and thickening properties. At the same time, pectin is also a natural water-soluble dietary fiber, which can reduce the risk of cancer, diabetes, and obesity, regulate the microenvironment of the human intestinal tract, and reduce blood lipids. High-ester pectin refers to pectin with a degree of esterification (the ratio of methyl-esterified galacturonic acid units to the total galacturonic acid units in pectin) ≥ 50%, because its morphological structure is closest to natural pectin molecules , has a good application prospect. The degree of esterification will affect the structure and functional properties of pectin and the ability to form complexes with other substances. It is important to keep the complete structure of pectin as much as possible and improve the performance of pectin for better application in food and other industries. significance.

果胶主要结构域包括同型半乳糖醛酸聚糖(HG)、鼠李半乳糖醛酸聚糖I(RG-I)、鼠李半乳糖醛酸聚糖II(RG-II),此外还有少量的取代型半乳糖醛酸聚糖。商品果胶主要是HG区域组成,常被用作食品相关领域的增稠剂和凝胶剂。利用高温强酸可以去除大部分RG-I区域提高果胶的稳定性和凝胶特性。而越来越多的研究表明,富含RG-I的果胶具有更好的抗癌功效、调节慢性代谢性疾病、以及免疫活性和益生元特性等。因此如何科学地提取具有完整RG-I果胶受到越来越多研究人员和消费者的关注。The main structural domains of pectin include homogalacturonan (HG), rhamnogalacturonan I (RG-I), rhamnogalacturonan II (RG-II), and A small amount of substituted galacturonan. Commercial pectin is mainly composed of HG domains and is often used as a thickener and gelling agent in food-related fields. Using high-temperature strong acid can remove most of the RG-I region to improve the stability and gel properties of pectin. More and more studies have shown that RG-I-rich pectin has better anti-cancer efficacy, regulation of chronic metabolic diseases, immune activity and prebiotic properties, etc. Therefore, how to scientifically extract pectin with complete RG-I has attracted the attention of more and more researchers and consumers.

现有的果胶提取方法主要包括离子交换法、酸/碱提取法、超临界萃取法和生物酶法等,但这些提取方法效果均不理想。此外强酸或强碱不仅对设备具有腐蚀性,提取后的废水还会造成环境污染。因此,如何制备出具有完整结构(RG-I)、性能好、产量高的高酯果胶是目前需要解决的问题之一。The existing pectin extraction methods mainly include ion exchange method, acid/alkali extraction method, supercritical extraction method and biological enzyme method, etc., but the effect of these extraction methods is not ideal. In addition, strong acid or strong alkali is not only corrosive to equipment, but also the extracted wastewater will cause environmental pollution. Therefore, how to prepare high-ester pectin with complete structure (RG-I), good performance and high yield is one of the problems to be solved at present.

发明内容Contents of the invention

本发明的目的是提供一种富含RG-I高酯的费约果果胶的提取方法,可以获得富含RG-I结构、品质好、产量高的费约果高酯果胶。The purpose of the present invention is to provide a method for extracting feijoa pectin rich in RG-I high ester, which can obtain feijoa high ester pectin rich in RG-I structure, good quality and high yield.

为达上述目的,本发明提供了一种富含RG-I高酯的费约果果胶的提取方法,包括以下步骤:For reaching above-mentioned object, the present invention provides a kind of extraction method of the feijoa pectin that is rich in RG-1 high ester, comprises the following steps:

(1)将费约果果肉煮沸后烘干至含水量低于6%,粉碎后将粉末与缓冲液混合均匀;(1) Boiling the feijoa pulp and drying it until the water content is lower than 6%, and mixing the powder and the buffer evenly after crushing;

(2)将混合后的混合液微波加热处理后依次酶解、离心、提纯后,将果胶干燥后即可;(2) Microwave heat treatment of the mixed mixed liquid followed by enzymatic hydrolysis, centrifugation and purification, followed by drying the pectin;

微波加热处理的功率为350-400W,时间为5-15min。The power of the microwave heating treatment is 350-400W, and the time is 5-15min.

进一步地,步骤(1)煮沸的温度为85-100℃,煮沸的时间为3-8min。Further, the boiling temperature in step (1) is 85-100° C., and the boiling time is 3-8 minutes.

进一步地,步骤(1)烘干的温度为45-55℃。Further, the drying temperature in step (1) is 45-55°C.

进一步地,步骤(1)粉碎后的粉末过60目筛。Further, the pulverized powder in step (1) is passed through a 60-mesh sieve.

进一步地,步骤(1)粉末与缓冲液的比例关系为10g:120-180mL,缓冲液为pH为6.8的50mM的柠檬酸钠缓冲液。Further, the ratio of powder to buffer in step (1) is 10 g: 120-180 mL, and the buffer is 50 mM sodium citrate buffer with a pH of 6.8.

进一步地,酶解过程具体为:将β-葡萄糖苷酶与费约果粉末按照250U/g的量混合均匀后,酶解处理90-100min,酶解的温度为45-55℃,酶解的转速为120-180rpm。Further, the enzymatic hydrolysis process is as follows: after mixing the β-glucosidase and feijoa powder uniformly according to the amount of 250U/g, enzymolysis treatment for 90-100min, the enzymolysis temperature is 45-55°C, and the enzymolysis The speed is 120-180rpm.

进一步地,步骤(2)酶解后,将提取后的溶液于90-100℃水浴环境中加热3-5min。Further, after the enzymatic hydrolysis in step (2), the extracted solution is heated in a water bath environment at 90-100° C. for 3-5 minutes.

进一步地,步骤(2)离心的转速为4500-5500rpm,离心的时间为8-12min。Further, the centrifugation speed in step (2) is 4500-5500 rpm, and the centrifugation time is 8-12 minutes.

进一步地,提纯具体包括以下步骤:收集离心后的上清液,向上清液中加入两倍体积的无水乙醇,混匀后静置沉淀2h,果胶析出后以4500-5500rpm进行离心,并用无水乙醇对试管中的果胶进行2-3次清洗。Further, the purification specifically includes the following steps: collect the supernatant after centrifugation, add twice the volume of absolute ethanol to the supernatant, mix well and let it settle for 2 hours, centrifuge at 4500-5500rpm after the pectin is precipitated, and use Clean the pectin in the test tube 2-3 times with absolute ethanol.

进一步地,果胶干燥的温度为45-60℃。Further, the drying temperature of pectin is 45-60°C.

综上所述,本发明具有以下优点:In summary, the present invention has the following advantages:

1、本发明公开的提取方法可以通过微波辅助酶解将费约果果肉中果胶较充分的提取出来,缩短了酶处理时间,大幅提高了提取效率和产品得率;而且本发明制备的果胶为高酯化度(酯化度68.43%)果胶,结构更加完整,RGΙ含量为41.38%,粘度较高。1. The extraction method disclosed in the present invention can fully extract the pectin in the feijoa pulp through microwave-assisted enzymolysis, shorten the enzyme treatment time, and greatly improve the extraction efficiency and product yield; and the fruit prepared by the present invention The gum is pectin with a high degree of esterification (68.43%), with a more complete structure, an RGI content of 41.38%, and a higher viscosity.

2、本发明公开的提取方法能耗低、污染小,提高了费约果的利用价值并为费约果的进一步开发利用提供理论指导。2. The extraction method disclosed by the present invention has low energy consumption and little pollution, improves the utilization value of feijoa fruit and provides theoretical guidance for further development and utilization of feijoa fruit.

3、本发明的方法更多地保留了果胶的RG-I区域结构,提取出了富含RG-I的高酯果胶,其中通过微波处理费约果果肉后再用生物酶可以将果胶从原料中分离出来,更多的原果胶转化为可溶性果胶被提取,因此本发明公开的提取方法可以得到高产量的果胶。3, the method of the present invention has kept the RG-I regional structure of pectin more, has extracted the high-ester pectin that is rich in RG-I, wherein can use biological enzyme again after the feijoa pulp by microwave treatment The gum is separated from the raw material, and more protopectin is converted into soluble pectin to be extracted, so the extraction method disclosed in the present invention can obtain high-yield pectin.

附图说明Description of drawings

图1为微波辅助酶法(MEHE)、酶法(EHE)和常规热酸水法(CHE)得到的果胶的稳定剪切流动曲线。Figure 1 shows the steady shear flow curves of pectin obtained by microwave-assisted enzymatic method (MEHE), enzymatic method (EHE) and conventional hot acid water method (CHE).

具体实施方式Detailed ways

以下结合实施例对本发明的原理和特征进行描述,所举实例只用于解释本发明,并非用于限定本发明的范围。实施例中未注明具体条件者,按照常规条件或制造商建议的条件进行。所用试剂或仪器未注明生产厂商者,均为可以通过市售购买获得的常规产品。The principles and features of the present invention are described below in conjunction with the examples, which are only used to explain the present invention, and are not intended to limit the scope of the present invention. Those who do not indicate the specific conditions in the examples are carried out according to the conventional conditions or the conditions suggested by the manufacturer. The reagents or instruments used were not indicated by the manufacturer, and they were all conventional products that could be purchased from the market.

本发明中采用的微波加热方法为使用微波炉加热方法,使用的微波炉型号为MM823ESJ-PA美的微波炉,加热时的档位为中低火。因此本发明在此同时附上对该中低火档位的频率的检测方法,以避免加热参数不清楚的问题。检测方法为:采用IEC705法对微波功率进行检测。首先,准备两个500mL的塑料烧杯并装满500mL温度为10℃左右的水,将烧杯放在烹饪区域的中央,将微波炉设置为全功率并打开电源,让微波炉精确运行60s,磁管控达到工作温度2s。然后从微波炉中取出烧杯,在用温度计测量每个烧杯的温度之前立即用磁力搅拌器搅拌每个烧杯并快速测温,每组重复两次取平均值。The microwave heating method that adopts in the present invention is to use microwave oven heating method, and the microwave oven model that uses is MM823ESJ-PA Midea microwave oven, and the stall during heating is low fire. Therefore, the present invention also adds a detection method for the frequency of the middle and low fire gears, so as to avoid the problem of unclear heating parameters. The detection method is: use IEC705 method to detect the microwave power. First, prepare two 500mL plastic beakers and fill them with 500mL of water at a temperature of about 10°C, place the beakers in the center of the cooking area, set the microwave oven to full power and turn on the power, let the microwave oven run precisely for 60s, and the magnetic control can work Temperature 2s. The beakers were then removed from the microwave oven, and each beaker was stirred with a magnetic stirrer immediately before the temperature of each beaker was measured with a thermometer, and the temperature was measured quickly, and each group was repeated twice to obtain an average value.

实施例1Example 1

本实施例提供了一种富含RG-I高酯的费约果果胶的提取方法,包括以下步骤:The present embodiment provides a kind of extraction method that is rich in the feijoa pectin of RG-1 high ester, comprises the following steps:

(1)对采摘回来的费约果洗涤擦干后去皮切分,随后放入90℃的去离子水中并煮沸5min以使其果胶酶失活。适当控水后置于50℃鼓风干燥箱中烘干至含水量小于6%,用粉碎机打粉后过60目筛制成费约果果肉粉(果肉和果浆)并将其密封于自封袋中,保存在干燥器中备用。(1) The plucked feijoas were washed, dried, peeled and cut into pieces, then placed in deionized water at 90° C. and boiled for 5 minutes to inactivate pectinase. After properly controlling the water, put it in a blast drying oven at 50°C and dry it until the water content is less than 6%. After pulverizing with a pulverizer, pass through a 60-mesh sieve to make feijoa pulp powder (pulp and pulp) and seal it in a self-sealing container. bag, and store in a desiccator for later use.

(2)取步骤(1)的费约果粉末10g于250mL三角瓶中,加入150mL 50mM柠檬酸钠缓冲液(料液比1:15),用柠檬酸调整pH为6.8,混合均匀。(2) Take 10g of feijoa powder from step (1) in a 250mL conical flask, add 150mL of 50mM sodium citrate buffer (solid-to-liquid ratio 1:15), adjust the pH to 6.8 with citric acid, and mix well.

(3)将(2)的混合液放入微波炉(MM823ESJ-PA美的微波炉)中,设定火候为中低火(380W IEC)处理9min后取出冷却至不烫手。(3) Put the mixture of (2) into a microwave oven (MM823ESJ-PA Midea microwave oven), set the heat to medium-low heat (380W IEC) for 9 minutes, take it out and cool it until it is not hot.

(4)称取β-葡萄糖苷酶(250U/g,酶活与底物的量比)于三角瓶中,混合均匀,在恒温震荡培养箱中进行酶解处理99min,酶解温度设定为50℃,摇床转速为150rpm。(4) Weigh β-glucosidase (250U/g, the ratio of enzyme activity to substrate) in a conical flask, mix evenly, and carry out enzymolysis treatment in a constant temperature shaking incubator for 99min, and the enzymolysis temperature is set at 50°C, the rotating speed of the shaker is 150rpm.

(5)提取结束后,在95℃水浴中加热5分钟使酶失活。(5) After the extraction, heat in a 95° C. water bath for 5 minutes to inactivate the enzyme.

(6)将提取后的溶液5000rpm进行离心10min,将上清液收集,并用蒸馏水对渣进行两次清洗,合并上清液后记录体积。(6) Centrifuge the extracted solution at 5000 rpm for 10 min, collect the supernatant, wash the residue twice with distilled water, and record the volume after combining the supernatant.

(7)向上清液中加入两倍体积的无水乙醇,混匀后静置沉淀两小时,果胶析出后5000rpm进行离心,并用无水乙醇对试管中的果胶进行2次清洗。(7) Add twice the volume of absolute ethanol to the supernatant, mix well and let it settle for two hours, centrifuge at 5000 rpm after the pectin is precipitated, and wash the pectin in the test tube twice with absolute ethanol.

(8)将清洗干净的果胶在电热鼓风干燥箱中,设定温度为50℃下烘干后,用研钵研磨后过100目筛备用。(8) Dry the cleaned pectin in an electric blast drying oven at a set temperature of 50° C., grind it with a mortar and pass it through a 100-mesh sieve for later use.

实施例2Example 2

(1)对采摘回来的费约果洗涤擦干后去皮切分,随后放入95℃的去离子水中并煮沸5min以使其果胶酶失活。适当控水后置于55℃鼓风干燥箱中烘干至含水量小于6%,用粉碎机打粉后过60目筛制成费约果果肉粉(果肉和果浆)并将其密封于自封袋中,保存在干燥器中备用。(1) Wash and dry the picked feijoas, peel and cut them into pieces, then put them into deionized water at 95° C. and boil them for 5 minutes to inactivate pectinase. After properly controlling the water, put it in a blast drying oven at 55°C and dry it until the water content is less than 6%. After pulverizing with a pulverizer, pass through a 60-mesh sieve to make feijoa pulp powder (pulp and pulp) and seal it in a self-sealing container. bag, and store in a desiccator for later use.

(2)取步骤(1)的费约果粉末10g于250mL三角瓶中,加入160mL 50mM柠檬酸钠缓冲液(料液比1:16),用柠檬酸调整pH为6.8,混合均匀。(2) Take 10g of feijoa powder from step (1) in a 250mL conical flask, add 160mL of 50mM sodium citrate buffer (solid-to-liquid ratio 1:16), adjust the pH to 6.8 with citric acid, and mix well.

(3)将(2)的混合液放入微波炉(MM823ESJ-PA美的微波炉)中,设定火候为中低火(350W IEC)处理10min后取出冷却至不烫手。(3) Put the mixture of (2) into a microwave oven (MM823ESJ-PA Midea microwave oven), set the heat to medium-low heat (350W IEC) for 10 minutes, take it out and cool it until it is not hot.

(4)称取β-葡萄糖苷酶(250U/g,酶活与底物的量比)于三角瓶中,混合均匀,在恒温震荡培养箱中进行酶解处理90min,酶解温度设定为50℃,摇床转速为150rpm。(4) Weigh β-glucosidase (250U/g, the ratio of enzyme activity to substrate) in a conical flask, mix well, and carry out enzymolysis treatment in a constant temperature shaking incubator for 90min, and the enzymolysis temperature is set at 50°C, the rotating speed of the shaker is 150rpm.

(5)提取结束后,在95℃水浴中加热5分钟使酶失活。(5) After the extraction, heat in a 95° C. water bath for 5 minutes to inactivate the enzyme.

(6)将提取后的溶液5500rpm进行离心8min,将上清液收集,并用蒸馏水对渣进行两次清洗,合并上清液后记录体积。(6) Centrifuge the extracted solution at 5500 rpm for 8 minutes, collect the supernatant, wash the residue twice with distilled water, and record the volume after combining the supernatant.

(7)向上清液中加入两倍体积的无水乙醇,混匀后静置沉淀两小时,果胶析出后5000rpm进行离心,并用无水乙醇对试管中的果胶进行3次清洗。(7) Add twice the volume of absolute ethanol to the supernatant, mix well and let it settle for two hours, centrifuge at 5000 rpm after the pectin is precipitated, and wash the pectin in the test tube 3 times with absolute ethanol.

(8)将清洗干净的果胶在电热鼓风干燥箱中,设定温度为50℃下烘干后,用研钵研磨后过100目筛备用。(8) Dry the cleaned pectin in an electric blast drying oven at a set temperature of 50° C., grind it with a mortar and pass it through a 100-mesh sieve for later use.

实施例3Example 3

(1)对采摘回来的费约果洗涤擦干后去皮切分,随后放入90℃的去离子水中并煮沸5min以使其果胶酶失活。适当控水后置于50℃鼓风干燥箱中烘干至含水量小于6%,用粉碎机打粉后过60目筛制成费约果果肉粉(果肉和果浆)并将其密封于自封袋中,保存在干燥器中备用。(1) The plucked feijoas were washed, dried, peeled and cut into pieces, then placed in deionized water at 90° C. and boiled for 5 minutes to inactivate pectinase. After properly controlling the water, put it in a blast drying oven at 50°C and dry it until the water content is less than 6%. After pulverizing with a pulverizer, pass through a 60-mesh sieve to make feijoa pulp powder (pulp and pulp) and seal it in a self-sealing container. bag, and store in a desiccator for later use.

(2)取步骤(1)的费约果粉末10g于250mL三角瓶中,加入140mL 50mM柠檬酸钠缓冲液(料液比1:14),用柠檬酸调整pH为6.8,混合均匀。(2) Take 10g of feijoa powder from step (1) in a 250mL conical flask, add 140mL of 50mM sodium citrate buffer (solid-to-liquid ratio 1:14), adjust the pH to 6.8 with citric acid, and mix well.

(3)将(2)的混合液放入微波炉(MM823ESJ-PA美的微波炉)中,设定火候为中低火(400W IEC)处理9min后取出冷却至不烫手。(3) Put the mixture of (2) into a microwave oven (MM823ESJ-PA Midea microwave oven), set the heat to medium-low heat (400W IEC) for 9 minutes, take it out and cool it until it is not hot.

(4)称取β-葡萄糖苷酶(250U/g,酶活与底物的量比)于三角瓶中,混合均匀,在恒温震荡培养箱中进行酶解处理100min,酶解温度设定为45℃,摇床转速为150rpm。(4) Weigh β-glucosidase (250U/g, the ratio of enzyme activity to substrate) in a conical flask, mix evenly, and carry out enzymolysis treatment in a constant temperature shaking incubator for 100min, and the enzymolysis temperature is set at 45°C, the rotation speed of the shaker is 150rpm.

(5)提取结束后,在90℃水浴中加热5分钟使酶失活。(5) After the extraction, heat in a 90° C. water bath for 5 minutes to inactivate the enzyme.

(6)将提取后的溶液5000rpm进行离心10min,将上清液收集,并用蒸馏水对渣进行两次清洗,合并上清液后记录体积。(6) Centrifuge the extracted solution at 5000 rpm for 10 min, collect the supernatant, wash the residue twice with distilled water, and record the volume after combining the supernatant.

(7)向上清液中加入两倍体积的无水乙醇,混匀后静置沉淀两小时,果胶析出后5500rpm进行离心,并用无水乙醇对试管中的果胶进行3次清洗。(7) Add twice the volume of absolute ethanol to the supernatant, mix well and let it settle for two hours, centrifuge at 5500 rpm after the pectin is precipitated, and wash the pectin in the test tube 3 times with absolute ethanol.

(8)将清洗干净的果胶在电热鼓风干燥箱中,设定温度为50℃下烘干后,用研钵研磨后过100目筛备用。(8) Dry the cleaned pectin in an electric blast drying oven at a set temperature of 50° C., grind it with a mortar and pass it through a 100-mesh sieve for later use.

对比例1Comparative example 1

本对比例提供了一种生物酶法提取费约果果胶的方法,包括以下步骤:This comparative example provides a kind of method that biological enzymatic method extracts feijoa pectin, comprises the following steps:

准确称取10g费约果果肉粉于锥形瓶中,加入150mL(1:15)pH为6.8的50mM柠檬酸钠溶液,再加入β-葡萄糖苷酶(250U/g)酶解99min,酶解温度设定为50℃,摇床转速为150rpm。其他处理与实施例1相同。Accurately weigh 10g of feijoa pulp powder into an Erlenmeyer flask, add 150mL (1:15) of 50mM sodium citrate solution with a pH of 6.8, then add β-glucosidase (250U/g) for enzymatic hydrolysis for 99min, and enzymatic hydrolysis The temperature was set at 50° C., and the rotation speed of the shaker was 150 rpm. Other treatments are the same as in Example 1.

对比例2Comparative example 2

本对比例提供了一种常规热酸水法提取费约果果胶的方法,包括以下步骤:This comparative example provides a kind of method that conventional hot acid water method extracts feijoa pectin, comprises the following steps:

(1)称取10g费约果果肉粉于烧杯中,加入150mL pH为1的盐酸溶液,在设定温度为95℃的集热式磁力搅拌器中提取130min。(1) Weigh 10 g of feijoa pulp powder into a beaker, add 150 mL of hydrochloric acid solution with a pH of 1, and extract in a heat-collecting magnetic stirrer with a set temperature of 95 °C for 130 min.

(2)提取完毕后使用离心机将果肉渣与可溶性化合物分离开来,分离得到的果肉渣用蒸馏水洗涤。(2) After the extraction is completed, a centrifuge is used to separate the pulp residue from the soluble compounds, and the separated pulp residue is washed with distilled water.

(3)收集滤液,在5000rpm的条件下,离心10min以除去细粉,加入2倍体积无水乙醇沉淀果胶2小时。(3) Collect the filtrate, centrifuge at 5000 rpm for 10 min to remove fine powder, add 2 times volume of absolute ethanol to precipitate pectin for 2 hours.

(4)沉淀的果胶在5000rpm的条件下离心10min后用无水乙醇洗涤果胶3次并再次离心,最终得到的果胶用电热鼓风干燥箱在50℃条件下干燥。(4) The precipitated pectin was centrifuged at 5000rpm for 10min, washed with absolute ethanol for 3 times and centrifuged again, and the finally obtained pectin was dried at 50°C in an electric blast drying oven.

试验例1-单糖测定Test Example 1-Monosaccharide Determination

单糖的测定方法采用三氯乙酸(TFA)法,精确称量多糖样品10mg,加入1mLTFA(4M)溶液在110℃下水解4小时。随后,水解产物在减压下与乙醇共蒸馏几次以完全除去TFA。通过添加盐酸羟胺、吡啶和乙酸酐混合物使水解样品和标准单糖乙酰化。使用日本岛津GC/MS气相色谱(QP 2010Plus)系统进行分析。初始温度设定为150℃并保持17min,温度程序以1℃/min的速度从150℃升至165℃,然后以10℃/min的速率升至200℃,再以50℃/mi的升温速度升至380℃。样品进样量1μL,分流比1:20,载气为氦气,流速1mL/min;离子源温度230℃,电子碰撞电压70eV,质量扫描范围m/z20-650,频率0.2s/scan,溶剂延迟时间设为5.5min。以β-苯基-葡萄糖苷(0.05%w/v)为内标进行标准定量,RG-I含量按照以下公式计算:The determination method of monosaccharide adopts trichloroacetic acid (TFA) method, accurately weighs 10mg of polysaccharide sample, adds 1mL TFA (4M) solution and hydrolyzes at 110 ℃ for 4 hours. Subsequently, the hydrolyzate was co-distilled several times with ethanol under reduced pressure to completely remove TFA. Hydrolyzed samples and standard monosaccharides were acetylated by adding a mixture of hydroxylamine hydrochloride, pyridine, and acetic anhydride. The Shimadzu GC/MS gas chromatography (QP 2010Plus) system was used for analysis. The initial temperature was set at 150°C and kept for 17 minutes. The temperature program increased from 150°C to 165°C at a rate of 1°C/min, then increased to 200°C at a rate of 10°C/min, and then increased at a rate of 50°C/min. Rise to 380°C. Sample injection volume 1μL, split ratio 1:20, carrier gas helium, flow rate 1mL/min; ion source temperature 230°C, electron impact voltage 70eV, mass scanning range m/z20-650, frequency 0.2s/scan, solvent The delay time is set to 5.5min. Standard quantification was performed with β-phenyl-glucoside (0.05% w/v) as the internal standard, and the RG-I content was calculated according to the following formula:

RG-I(%)=2Rha(%)+Ara(%)+Gal(%)RG-I(%)=2Rha(%)+Ara(%)+Gal(%)

试验例2-酯化度(DE)测定Test example 2-determination of degree of esterification (DE)

酯化度的测定方法参照食品安全国家标准GB25533 2010中的滴定法进行。The determination method of the degree of esterification is carried out with reference to the titration method in the national food safety standard GB25533 2010.

试验例3-流变学特性测定Test Example 3 - Determination of Rheological Properties

将果胶样品溶于去离子水中配制1.0%(w/v)的浓度。旋转流变仪为美国Thermo公司旗下的HaakeRS6000,探头使用平面圆形钢板(直径60mm),间隙设置1.0mm,测定样品的表观粘度。在25℃下测量随剪切速率从0.1s-1逐渐增加至100s-1下的表观粘度曲线。Pectin samples were dissolved in deionized water to make a concentration of 1.0% (w/v). The rotational rheometer is a Haake RS6000 owned by American Thermo Company. The probe uses a flat circular steel plate (diameter 60 mm), and the gap is set at 1.0 mm to measure the apparent viscosity of the sample. The apparent viscosity curve was measured at 25°C with the shear rate gradually increasing from 0.1 s -1 to 100 s -1 .

实验结果如表1和图1所示。The experimental results are shown in Table 1 and Figure 1.

表1果胶理化参数Table 1 Physical and chemical parameters of pectin

Figure BDA0004039401110000081
Figure BDA0004039401110000081

由表1和图1可知,采用本发明公开的提取方法,可以大幅度提高果胶的得率,并且在酯化度和RG-Ⅰ含量均保持较高的水平。本发明公开的提取方法可以更多地保留果胶的RG-I区域结构,提取出富含RG-I的高酯果胶。同时本发明提取出的果胶酯化度为68.43%,属于高酯果胶,且粘度与酶法提取的果胶相当,优于常规热酸水法提取果胶。It can be seen from Table 1 and Figure 1 that the extraction method disclosed in the present invention can greatly increase the yield of pectin, and maintain a relatively high level of esterification degree and RG-I content. The extraction method disclosed by the invention can retain more RG-I domain structure of pectin, and extract high-ester pectin rich in RG-I. At the same time, the degree of esterification of pectin extracted by the invention is 68.43%, which belongs to high-ester pectin, and its viscosity is equivalent to that of pectin extracted by enzymatic method, which is better than that of pectin extracted by conventional hot acid water method.

虽然对本发明的具体实施方式进行了详细地描述,但不应理解为对本专利的保护范围的限定。在权利要求书所描述的范围内,本领域技术人员不经创造性劳动即可作出的各种修改和变形仍属本专利的保护范围。Although the specific implementation of the present invention has been described in detail, it should not be construed as limiting the protection scope of this patent. Within the scope described in the claims, various modifications and deformations that can be made by those skilled in the art without creative work still belong to the protection scope of this patent.

Claims (10)

1.一种富含RG-I高酯的费约果果胶的提取方法,其特征在于,包括以下步骤:1. an extraction method of feijoa pectin rich in RG-1 high ester, is characterized in that, comprises the following steps: (1)将费约果果肉煮沸后烘干至含水量低于6%,粉碎后将粉末与缓冲液混合均匀;(1) Boiling the feijoa pulp and drying it until the water content is lower than 6%, and mixing the powder and the buffer evenly after crushing; (2)将混合后的混合液微波加热处理后依次酶解、离心、提纯后,将果胶干燥后即可;(2) Microwave heat treatment of the mixed mixed liquid followed by enzymatic hydrolysis, centrifugation and purification, followed by drying the pectin; 所述微波加热处理的功率为350-400W,时间为5-15min。The power of the microwave heating treatment is 350-400W, and the time is 5-15min. 2.如权利要求1所述的富含RG-I高酯的费约果果胶的提取方法,其特征在于,所述步骤(1)煮沸的温度为85-100℃,煮沸的时间为3-8min。2. the extraction method of the feijoa pectin that is rich in RG-1 high ester as claimed in claim 1, is characterized in that, the temperature of described step (1) boiling is 85-100 ℃, and the time of boiling is 3 -8min. 3.如权利要求1所述的富含RG-I高酯的费约果果胶的提取方法,其特征在于,所述步骤(1)烘干的温度为45-55℃。3. the extraction method of the feijoa pectin that is rich in RG-1 high ester as claimed in claim 1, is characterized in that, the temperature of described step (1) drying is 45-55 ℃. 4.如权利要求1所述的富含RG-I高酯的费约果果胶的提取方法,其特征在于,所述步骤(1)粉碎后的粉末过60目筛。4. the extraction method of the feijoa pectin that is rich in RG-1 high ester as claimed in claim 1, is characterized in that, the powder after described step (1) pulverizes crosses 60 mesh sieves. 5.如权利要求1或4所述的富含RG-I高酯的费约果果胶的提取方法,其特征在于,所述步骤(1)粉末与缓冲液的比例关系为10g:120-180mL,所述缓冲液为pH为6.8的50mM的柠檬酸钠缓冲液。5. the extraction method of the feijoa pectin that is rich in RG-1 high ester as claimed in claim 1 or 4 is characterized in that, the ratio relation of described step (1) powder and damping fluid is 10g: 120- 180 mL, and the buffer solution is a 50 mM sodium citrate buffer solution with a pH of 6.8. 6.如权利要求1所述的富含RG-I高酯的费约果果胶的提取方法,其特征在于,所述酶解过程具体为:将β-葡萄糖苷酶与费约果粉末按照250U/g的量混合均匀后,酶解处理90-100min,酶解的温度为45-55℃,酶解的转速为120-180rpm。6. the extraction method of the feijoa pectin that is rich in RG-1 high ester as claimed in claim 1, is characterized in that, described enzymolysis process is specially: with β-glucosidase and feijoa powder according to After the amount of 250U/g is mixed evenly, the enzymolysis treatment is carried out for 90-100min, the temperature of the enzymolysis is 45-55°C, and the rotation speed of the enzymolysis is 120-180rpm. 7.如权利要求1所述的富含RG-I高酯的费约果果胶的提取方法,其特征在于,所述步骤(2)酶解后,将提取后的溶液于90-100℃水浴环境中加热3-5min。7. the extraction method of the feijoa pectin that is rich in RG-1 high ester as claimed in claim 1, is characterized in that, after described step (2) enzymolysis, the solution after extraction is at 90-100 ℃ Heat in a water bath environment for 3-5min. 8.如权利要求1所述的富含RG-I高酯的费约果果胶的提取方法,其特征在于,所述步骤(2)离心的转速为4500-5500rpm,离心的时间为8-12min。8. the extraction method of the feijoa pectin that is rich in RG-1 high ester as claimed in claim 1, is characterized in that, the centrifugal rotating speed of described step (2) is 4500-5500rpm, and the time of centrifugal is 8- 12min. 9.如权利要求1所述的富含RG-I高酯的费约果果胶的提取方法,其特征在于,所述提纯具体包括以下步骤:收集离心后的上清液,向上清液中加入两倍体积的无水乙醇,混匀后静置沉淀2h,果胶析出后以4500-5500rpm进行离心,并用无水乙醇对试管中的果胶进行2-3次清洗。9. the extraction method of the feijoa pectin that is rich in RG-1 high ester as claimed in claim 1, is characterized in that, described purification specifically comprises the following steps: collect the supernatant after centrifugation, in supernatant Add twice the volume of absolute ethanol, mix well and let it settle for 2 hours, centrifuge at 4500-5500rpm after the pectin is precipitated, and wash the pectin in the test tube 2-3 times with absolute ethanol. 10.如权利要求1所述的富含RG-I高酯的费约果果胶的提取方法,其特征在于,所述果胶干燥的温度为45-60℃。10. the method for extracting the feijoa pectin that is rich in RG-1 high ester as claimed in claim 1, is characterized in that, the temperature of described pectin drying is 45-60 ℃.
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