CN111518854A - 一种魔芋精粉多酶酶解物及其制备方法 - Google Patents
一种魔芋精粉多酶酶解物及其制备方法 Download PDFInfo
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- CN111518854A CN111518854A CN202010248586.9A CN202010248586A CN111518854A CN 111518854 A CN111518854 A CN 111518854A CN 202010248586 A CN202010248586 A CN 202010248586A CN 111518854 A CN111518854 A CN 111518854A
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- konjac
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- zymolyte
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Abstract
本发明提供了一种魔芋精粉多酶酶解物及其制备方法,属于功能食品加工领域。本发明通过将魔芋精粉原料在酒精中纯化,得到魔芋湿精粉;再将魔芋湿精粉进行脱醇处理得到纯化魔芋精粉;再加水和酶进行酶解,最终得到魔芋精粉多酶酶解物。本发明通过对工艺进行优化,得到一种含有一定速溶性多糖和低聚糖的酶解产物。
Description
技术领域
本发明涉及功能食品加工技术领域,尤其涉及一种魔芋精粉多酶酶解物及其制备方法。
背景技术
魔芋有较高的功能和医学价值,现代研究表明魔芋块茎的主要化学成分为:葡甘聚糖、淀粉、其他多糖、粗蛋白、必需氨基酸,以及钾、钙、镁、钠、铁、锰、铜等人体必需的多种微量元素。
魔芋中的葡甘聚糖是继淀粉和纤维素之后,一种较为丰富的可再生天然高分子资源,具有可生物降解性和水溶性,是自然界分子量较大、黏度最高的膳食纤维,它能吸附其自身体积200倍的水分形成粘稠的溶液,可增强饱腹感,因不被人体的消化酶所影响,而不会产生热量。是公认的可溶性多功能性多糖,且其适度降解后又可产生功能更强大的魔芋低聚糖。但其吸水溶胀、溶解速度很慢,通常在2小时以上。
目前魔芋葡甘聚糖酶解有以下可优化的问题:①单酶酶解致使用酶量大,酶解时间过长,且魔芋精粉利用率低。②有的酶解法是以检测还原糖为主要指标,还原糖量高不代表低聚糖含量高,相反在相同可溶性固形物含量的前提下,还原糖含量越高,低聚糖含量越低,则这种酶解工艺也失去了制备低聚糖的意义。③有的酶解法只能应用于实验室研究,无法投入正式生产。④魔芋精粉具有吸水膨胀的特性,未经降解直接食用后易产生腹胀感,存在一定安全风险;对其适度降解后既可消除这一缺陷,又可提高溶解速度,且保持原有的功能特性。
发明内容
本发明的目的在于提供一种魔芋精粉多酶酶解物及其制备方法,克服了现有技术中存在的酶解时间长、酶解不彻底、魔芋精粉利用率低等缺陷,得到一种含有一定速溶性多糖和低聚糖的酶解产物。
为了实现上述发明目的,本发明提供以下技术方案:
本发明提供了一种魔芋精粉多酶酶解物的制备方法,包括以下步骤:
(1)将魔芋精粉原料在酒精中纯化,得到魔芋湿精粉;
(2)将魔芋湿精粉进行脱醇处理,得到纯化魔芋精粉;
(3)将所得纯化魔芋精粉、水和酶混合后进行酶解,即得魔芋精粉多酶酶解物。
作为优选,所述步骤(1)中酒精的体积浓度为55%~75%;
所述步骤(1)中魔芋精粉原料与酒精的料液比为1g:3~5ml;
所述步骤(1)中纯化的时间为1~3h;
所述步骤(2)中脱醇处理的温度为50~70℃。
作为优选,所述步骤(3)中纯化魔芋精粉与水的质量体积比为15~30g:100mL;
所述步骤(3)中的酶包含葡甘聚糖酶、纤维素酶和果胶酶;
所述葡甘聚糖酶的用量为50~70u/g纯化魔芋精粉,所述纤维素酶的用量为40~60u/g纯化魔芋精粉,所述果胶酶占体系总质量的0.03%~0.05%;
所述酶解的温度为55~65℃,酶解的时间为2~3h。
本发明还提供了所述制备方法得到的魔芋精粉多酶酶解物。
本发明提供了一种魔芋精粉多酶酶解物及其制备方法,通过本方法得到的酶解物中功能成分(如低聚糖)含量为5.69g/100g~5.86g/100g精粉,占总糖含量的39.33%~40.5%,还含有一定量的速溶性多糖。该产品具有预防肥胖、降低血脂、预防肝脂肪变性的作用;具有降血糖,抗炎、提高机体清除自由基的作用,对糖尿病模型大鼠的高血糖具有改善作用。
本发明以魔芋精粉为原料,采用复合酶酶解,加速酶解过程,加大酶解效率,提高酶解产物品质。通过对工艺参数进行优化,得到一种在用酶选择、酶用量、酶解时间等生产工艺参数上有所优化且又含有一定速溶性多糖和低聚糖的酶解产物。
具体实施方式
本发明提供了一种魔芋精粉多酶酶解物的制备方法,包括以下步骤:
(1)将魔芋精粉原料在酒精中纯化,得到魔芋湿精粉;
(2)将魔芋湿精粉进行脱醇处理,得到纯化魔芋精粉;
(3)将所得纯化魔芋精粉、水和酶混合后进行酶解,即得魔芋精粉多酶酶解物。
在本发明中,所述魔芋精粉优选为高纯度市售的魔芋精粉。
在本发明中,所述步骤(1)中酒精的体积浓度优选为55%~75%,进一步优选为60%~70%,更进一步优选为62%~68%。
在本发明中,所述步骤(1)中魔芋精粉原料与酒精的料液比优选为1g:3~5ml,进一步优选为1g:3.5~4.5ml。
在本发明中,所述步骤(1)中纯化的时间优选为1~3h,进一步优选为1.5~2.5h。
在本发明中,所述纯化优选将魔芋精粉原料与酒精搅拌混匀并浸泡。
在本发明中,所述纯化是为了将魔芋精粉中含有的的腥臭味、麻辣味、SO2等除去。
在本发明中,所述脱醇处理优选热风干燥的手段进行。
在本发明中,所述脱醇处理的温度优选为50~70℃,进一步优选为55~65℃,更进一步优选为58~62℃。
在本发明中,所述脱醇处理是为了除去酒精味,以闻不到酒精味为准。
在本发明中,所述步骤(3)中纯化魔芋精粉与水的质量体积比优选为15~30g:100mL,进一步优选为20~25g:100mL。
在本发明中,所述步骤(3)中的酶优选包含葡甘聚糖酶、纤维素酶和果胶酶。
在本发明中,所述葡甘聚糖酶的用量优选为50~70u/g纯化魔芋精粉,进一步优选为55~65u/g纯化魔芋精粉,更进一步优选为60~62u/g纯化魔芋精粉;所述纤维素酶的用量优选为40~60u/g纯化魔芋精粉,进一步优选为45~55u/g纯化魔芋精粉,更进一步优选为47~52u/g纯化魔芋精粉;所述果胶酶的用量优选为占体系总质量的0.03%~0.05%,进一步优选为占体系总质量的0.035%~0.04%。
在本发明中,所述体系是指纯化魔芋精粉、酶和水混合得到的混合物。
在本发明中,所述酶解的温度优选为55~65℃,进一步优选为58~60℃;酶解的时间优选为2~3h。
本发明所述酶解的原理为:依据酶的专一性,同时用葡甘聚糖酶和纤维素酶将葡甘聚糖酶解为低聚合度魔芋多糖和魔芋低聚糖,果胶酶将果胶酶解为半乳糖醛酸以破坏魔芋精粉颗粒的结构,从而提高其它酶的作用效果。
本发明还提供了所述制备方法得到的魔芋精粉多酶酶解物。
下面结合实施例对本发明提供的技术方案进行详细的说明,但是不能把它们理解为对本发明保护范围的限定。
实施例1
步骤1:纯化:将魔芋精粉原料在体积浓度为55%的酒精中纯化,魔芋精粉原料与酒精的料液比为1g:3ml,纯化1h后得到魔芋湿精粉。
步骤2:脱醇:将魔芋湿精粉进行脱醇处理,脱醇的温度为50℃,脱醇后得到纯化魔芋精粉;
步骤3:酶解:取15g纯化魔芋精粉加100mL水,再加葡甘聚糖酶、纤维素酶和果胶酶酶解,其中葡甘聚糖酶的用量为50u/g纯化魔芋精粉,纤维素酶的用量为40u/g纯化魔芋精粉,果胶酶的用量占体系总质量的0.03%,在55℃条件下酶解2h即得魔芋精粉多酶酶解物。
所得酶解物中功能成分(低聚糖)含量为5.86g/100g精粉,占总糖含量的40.5%。
实施例2
步骤1:纯化:将魔芋精粉原料在体积浓度为60%的酒精中纯化,魔芋精粉原料与酒精的料液比为1g:4ml,纯化1.5h后得到魔芋湿精粉。
步骤2:脱醇:将魔芋湿精粉进行脱醇处理,脱醇的温度为65℃,脱醇后得到纯化魔芋精粉;
步骤3:酶解:取25g纯化魔芋精粉加100mL水,再加葡甘聚糖酶、纤维素酶和果胶酶酶解,其中葡甘聚糖酶的用量为60u/g纯化魔芋精粉,纤维素酶的用量为50u/g纯化魔芋精粉,果胶酶的用量占体系总质量的0.04%,在60℃条件下酶解2.5h即得魔芋精粉多酶酶解物。
所得酶解物中功能成分(低聚糖)含量为5.78g/100g精粉,占总糖含量的39.95%。
实施例3
步骤1:纯化:将魔芋精粉原料在体积浓度为75%的酒精中纯化,魔芋精粉原料与酒精的料液比为1g:5ml,纯化3h后得到魔芋湿精粉。
步骤2:脱醇:将魔芋湿精粉进行脱醇处理,脱醇的温度为70℃,脱醇后得到纯化魔芋精粉;
步骤3:酶解:取30g纯化魔芋精粉加100mL水,再加葡甘聚糖酶、纤维素酶和果胶酶酶解,其中葡甘聚糖酶的用量为70u/g纯化魔芋精粉,纤维素酶的用量为60u/g纯化魔芋精粉,果胶酶的用量占体系总质量的0.05%,在65℃条件下酶解3h即得魔芋精粉多酶酶解物。
所得酶解物中功能成分(低聚糖)含量为5.69g/100g精粉,占总糖含量的39.33%。
实施例4
对实施例1-3制备的魔芋精粉多酶酶解物进行动物实验。
步骤1:动物造模和分组
选择健康雌性小鼠50只,饲喂普通饲料2周后,将它们随机分为2组,一组为空白对照组10只(饲喂普通饲料)、另一组为模型对照组40只(饲喂高脂饲料),两组给予无限量饲料,喂养4周后,成功建立小鼠肥胖模型,模型对照组任意分为4组,每组小鼠10只,4组小鼠分别为高脂模型组(饲喂高脂饲料)、魔芋精粉酶解物低剂量组(饲喂高脂饲料并灌胃酶解物15mg·100g-1(BW)·d-1)、魔芋精粉酶解物中剂量组(饲喂高脂饲料并灌胃酶解物30mg·100g-1(BW)·d-1)、魔芋精粉酶解物高剂量组(饲喂高脂饲料并灌胃酶解物60mg·100g-1(BW)·d-1)。空白对照组继续用普通饲料饲喂小鼠,高脂模型组以及魔芋粉酶解物低、中、高剂量组继续给小鼠饲喂高脂饲料,连续灌胃魔芋粉酶解物一个月。饲养温度为24~27℃,相对湿度45%~50%,采用11h:13h昼夜间断照明。
步骤2:样品采集
喂养10周后,各组小鼠禁食12h,处死前称体质量,麻醉后快速摘眼球取血,颈椎脱臼致死,快速分离出肝脏、心脏、脾脏、肾脏、肺、肾周围脂肪和腹股沟皮下脂肪,用盐水冲洗器官,滤纸吸干,称重脏器和组织。
步骤3:指标测定
在实验期间每天记录小鼠生长状态(小鼠的毛色、进食、排便和活动),分别于周三、周四检测小鼠的日摄食量以及日饮水量,即于周三、周四称量饲料总量和加水总量,于周四、周五同一时间称量剩余饲料量和剩余水量。(即摄食量=饲料总量-剩余饲料量,饮水量=总水量-残留水量)。每周周六称量1次体质量。
切开腹部,取出小鼠体内的所有脂肪(包括肾周围的脂肪组织和腹股沟的皮下脂肪组织)并称量脂肪,按照公式(1)计算脂肪指数。
脂肪指数/%=m1/m×100 (1)
式中:m为小鼠处死前体质量/g;m1为小鼠体内各脂肪组织质量/g。
步骤4:数据处理
利用SPSS 20软件进行统计学分析,用均值±标准差表示,各组间比较采用单因素方差分析,P<0.05表示两组之间存在显著性差异。
表1魔芋酶解物对小鼠日饮水量影响/mL(均值±标准差)
注:#表示与高脂模型组相比在P<0.05有差异。
从表1可以看出,高脂模型组与魔芋酶解物低、中、高剂量组有显著差异(P<0.05),说明魔芋酶解物在一定程度上可降低小鼠饮水量,魔芋酶解物高剂量组效果最明显。
实施例5
对实施例1-3制备的魔芋精粉多酶酶解物进行动物实验。
步骤1:糖尿病大鼠模型的构建
选取60只健康雌性大鼠正常喂养1周,期间定期测量实验动物的饮水量、采食量。1周后,测量记录实验动物的体重及空腹血糖值,并进行随机分组(正常对照组和5组造模组),每组10只。造模组大鼠通过腹腔注射内含STZ的柠檬酸-柠檬酸钠缓冲液诱导糖尿病,(用预冷的柠檬酸盐缓冲液pH=4.5以1%的浓度溶解STZ,经0.22mol/L微孔滤膜过滤除菌。STZ容易失活,STZ快速称取后,用锡纸包裹避光,且现配现用并时刻置于冰浴中备用),注射STZ浓度为45mg/kg,2%注射量,正常组大鼠腹腔注射等体积的柠檬酸-柠檬酸钠缓冲液,注射STZ过程中需要对大鼠腹腔皮肤进行消毒,防止感染。注射结束后,各组继续维持原来饮食。24h后于大鼠尾部采血,测量其空腹血糖值,使用血糖仪及配套血糖试纸测量大鼠空腹血糖,当造模组大鼠空腹血糖>16.7mmol/L(可重复测量2-3次,血糖稳定),且出现有多饮多尿,体重减轻症状,视为建模成功。
步骤2:动物分组及给药
将造模成功的糖尿病大鼠进行分组调整,分别为魔芋精粉酶解物高、中、低剂量组、阳性药物组、糖尿病模型组,血糖值与造模前一致的设为正常组。连续灌胃给药6周,实验期间,各组大鼠正常自由饮水摄食。灌胃时间为每日下午4点,各组大鼠灌胃给药剂量如下:魔芋精粉酶解物高、中、低组灌胃剂量分别为2.500g/(kg·d)、1.250g/(kg·d)、0.625g/(kg·d),阳性药物组给药剂量为0.140g/(kg·d),正常组及高糖模型组饲喂等量生理盐水。
步骤3:指标测定
在大鼠灌胃给药期间每周测量各组大鼠的体重并记录,结果以g表示,分析并比较各组大鼠的体重差异;测定各组实验大鼠的空腹血糖值,结果表示为mmol/L,分析比较各组大鼠的血糖水平变化;测量并记录实验动物的饮水量、采食量,并分析灌胃给药期间各组大鼠饮水量和采食量的变化。
步骤4:统计学方法
利用SPSS20软件进行统计学分析,用均值±标准差表示,各组间比较采用单因素方差分析,P<0.05表示两组之间存在显著性差异。
表2魔芋酶解物对糖尿病大鼠饮水量的影响
注:模型组与正常组相比,**P<0.01,表示差异极显著;治疗组与模型组相比,##P<0.01,表示差异极显著;魔芋酶解物剂量组与阳性药物组相比,&&P<0.01,表示差异极显著。
由表2可看出,给药第0d,模型组饮水量较正常组极显著上升(p<0.01),给药21d后,阳性药物组及魔芋酶解物各剂量组饮水量均有不同程度下降,给药第42d,模型组饮水量较正常组具有极显著差异(p<0.01),阳性药物组及魔芋酶解物各剂量组饮水量较模型组具有极显著差异(p<0.01),且魔芋酶解物高、中剂量组饮水量与阳性药物组无显著差异。由此可知魔芋酶解物可缓解糖尿病大鼠多饮症状,且一定剂量的魔芋酶解物就糖尿病患者多饮症状的缓解具有与二甲双胍相似的效果。
由以上实施例可知,本发明提供了一种魔芋精粉多酶酶解物的制备方法,动物(白鼠)实验表明,该产品具有预防肥胖、降低血脂、预防肝脂肪变性的作用;具有降血糖,降低脂质过氧化、抗炎、提高机体清除自由基的作用,对糖尿病模型大鼠的高血糖具有改善作用。
以上所述仅是本发明的优选实施方式,应当指出,对于本技术领域的普通技术人员来说,在不脱离本发明原理的前提下,还可以做出若干改进和润饰,这些改进和润饰也应视为本发明的保护范围。
Claims (4)
1.一种魔芋精粉多酶酶解物的制备方法,其特征在于,包括以下步骤:
(1)将魔芋精粉原料在酒精中纯化,得到魔芋湿精粉;
(2)将魔芋湿精粉进行脱醇处理,得到纯化魔芋精粉;
(3)将所得纯化魔芋精粉、水和酶混合后进行酶解,即得魔芋精粉多酶酶解物。
2.如权利要求1所述的制备方法,其特征在于,所述步骤(1)中酒精的体积浓度为55%~75%;
所述步骤(1)中魔芋精粉原料与酒精的料液比为1g:3~5ml;
所述步骤(1)中纯化的时间为1~3h;
所述步骤(2)中脱醇处理的温度为50~70℃。
3.如权利要求1所述的制备方法,其特征在于,所述步骤(3)中纯化魔芋精粉与水的质量体积比为15~30g:100mL;
所述步骤(3)中的酶包含葡甘聚糖酶、纤维素酶和果胶酶;
所述葡甘聚糖酶的用量为50~70u/g纯化魔芋精粉,所述纤维素酶的用量为40~60u/g纯化魔芋精粉,所述果胶酶占体系总质量的0.03%~0.05%;
所述酶解的温度为55~65℃,酶解的时间为2~3h。
4.权利要求1~3任意一项所述制备方法得到的魔芋精粉多酶酶解物。
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