CN112920285B - 一种米糠多糖的制备方法及其应用 - Google Patents

一种米糠多糖的制备方法及其应用 Download PDF

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CN112920285B
CN112920285B CN202010811923.0A CN202010811923A CN112920285B CN 112920285 B CN112920285 B CN 112920285B CN 202010811923 A CN202010811923 A CN 202010811923A CN 112920285 B CN112920285 B CN 112920285B
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CN112920285A (zh
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刘玉春
郭伟群
庄绪会
陈园
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Abstract

本发明公开一种米糠多糖的制备方法及其应用。本发明米糠多糖的制备方法,包括:将米糠纤维浸入水中进行热水浸提和超声浸提,获得第一提取液;将第一提取液固液分离,获得第一固体沉淀和第一上清液,第一上清液干燥得第一米糠多糖;将第一固体沉淀的一部分进行烘干和粉碎后作为碳源发酵培养裂褶菌,获得发酵液,将发酵液固液分离后浓缩成复合酶制剂;用复合酶制剂处理剩余的第一固体沉淀,热水浸提和超声浸提,获得第二提取液;将第二提取液固液分离,获得第二上清液并干燥得第二米糠多糖;将第一米糠多糖和第二米糠多糖合并得到米糠多糖。本发明以米糠纤维为主要原料进行发酵和酶解制备的米糠多糖,产品得率高,绿色无污染,适用范围广。

Description

一种米糠多糖的制备方法及其应用
技术领域
本发明涉及多糖的制备工艺技术领域。更具体地,涉及一种米糠多糖的制备方法及其应用。
背景技术
中国稻谷产量达到2.1亿吨,米糠是稻谷脱壳精碾糙米后的主要副产物,稻谷加工过程出糠率在5%左右。米糠多糖具有抗氧化、抗肿瘤和提高免疫等多种生理活性。米糠到目前为止还没有有效地利用,大部分被用于饲料甚至废弃,提取米糠多糖对于增加稻谷附加值具有现实意义。米糠多糖的主流提取方法是热水浸提法,多次改良后报道了微波协同提取法、高压脉冲提取法、碱性条件浸提方法等,但得率提高并不明显或对环境会造成污染,不符合绿色环保的理念。因此,亟需一种得率高且环保的提取方法。
调味酱是用于协调各类食品的味道,以满足食用者要求的酱状调味品。高盐高油体系下,酱料体系存在油析、分层等问题,工业生产中通常加入稳定剂来进行增稠和乳化稳定,比如变性淀粉、黄原胶等乳化剂。米糠多糖作为食物纤维强化的用途很多,但是没有发现其作为调味酱乳化稳定剂来应用的研究。
发明内容
本发明的一个目的在于提供一种米糠多糖的制备方法,该方法得率高且环保。
本发明的另一个目的在于提供上述方法得到的米糠多糖在提高乳化能力中的应用及活性成分为米糠多糖的乳化剂。
为达到上述目的,本发明提供了如下技术方案:
本发明首先提供了一种米糠多糖的制备方法,包括如下步骤:
将米糠纤维浸入水中进行热水浸提和超声浸提,获得第一提取液;
将所述第一提取液进行固液分离,获得第一固体沉淀和第一上清液,对第一上清液进行干燥,得第一米糠多糖;
将第一固体沉淀的一部分进行烘干和粉碎后,作为发酵培养基的碳源发酵培养裂褶菌,获得发酵液,将所述发酵液进行固液分离后浓缩成复合酶制剂;
用所述复合酶制剂处理剩余的第一固体沉淀,再次进行所述热水浸提和超声浸提,获得第二提取液;
将所述第二提取液进行固液分离,获得第二上清液,对第二上清液进行干燥,得第二米糠多糖;
将第一米糠多糖和第二米糠多糖合并得到米糠多糖。
上述制备方法中,也可以将第一上清液和第二上清液合并后进行干燥得到米糠多糖。
上述制备方法中,所述米糠纤维与水的质量比(即料液比)为1:8~1:15(例如可以为1:8、1:9、1:10、1:11、1:12、1:13、1:14和1:15以及任意比例间的任意范围)。
上述制备方法中,所述热水浸提为在pH 3.0,温度为80℃~100℃(例如可以为80℃、85℃、90℃、95℃和100℃以及任意温度间的任意范围)的条件下浸提120min。
上述制备方法中,所述超声浸提是在温度为70℃~100℃(例如可以为70℃、75℃、80℃、85℃、90℃、95℃和100℃以及任意温度间的任意范围)、超声功率为150W~300W(例如可以为150W、200W、250W、280W和300W以及任意功率间的任意范围)的条件下浸提90min。
上述制备方法中,所述发酵培养基除了碳源之外还加入了无机盐(所述无机盐的组分及其占发酵培养基的质量体积百分比为0.02%尿素,0.42%(NH4)2SO4,0.2%KH2PO4,0.03%CaCl2,0.03%MgSO4·H2O,0.2%吐温80,和体积百分比为0.2%微量元素溶液(微量元素溶液的组分及其占微量元素溶液的质量体积百分比为0.5%FeSO4·7H2O,0.16%MnSO4·4H2O,0.14%ZnSO4·7H2O,0.2%CoCl2和余量的水))和余量的水,所述碳源占所述发酵培养基的质量体积百分比为1~10%(即所述碳源的质量/所述发酵培养基的体积×100=1~10%)。
上述制备方法中,所述发酵培养是在温度为30℃、转速为220rpm的培养条件下发酵培养2~10天。本发明发酵培养的时间是根据发酵过程中取样测定发酵液酶活(包括葡聚糖酶、木聚糖酶、α-L-阿拉伯呋喃糖苷酶、阿魏酸酯酶、α-葡糖醛酸酶)和蛋白含量确定的,优选的发酵培养是在温度为30℃、转速为220rpm的培养条件下发酵培养3天。
上述制备方法中,所述浓缩为浓缩2~10倍,具体为用截留分子量1000~10000Da中空纤维柱浓缩。
本发明中应用iTRAQ蛋白质组分析技术测定复合酶制剂的组成,包含40个家族(GH1、GH2、GH3、GH5、GH6、GH7、GH10、GH11、GH13、GH15、GH16、GH17、GH18、GH25、GH27、GH28、GH30、GH31、GH35、GH37、GH43、GH45、GH47、GH51、GH53、GH55、GH61、GH62、GH71、GH74、GH76、GH79、GH81、GH88、GH92、GH93、GH95、GH105、GH115、GH125)的75个糖苷水解酶蛋白;具体的,该复合酶制剂包含以下活性的一种或多种:内切-β-1,4-葡聚糖酶/纤维素酶(EC3.2.1.4)活性和/或内切-β-1,4-木聚糖酶(EC3.2.1.8)活性和/或β-葡糖苷酶(EC 3.2.1.21)活性和/或β-甘露糖苷酶(EC3.2.1.25)活性和/或葡聚糖β-1,3-葡糖苷酶(EC 3.2.1.58)活性和/或外切-β-1,4-葡聚糖酶/纤维糊精酶(EC 3.2.1.74)活性和/或葡聚糖内切-1,6-β-葡糖苷酶(EC3.2.1.75)活性和/或甘露聚糖内切-β-1,4-甘露糖苷酶(EC 3.2.1.78)活性和/或纤维素β-1,4-纤维二糖苷酶(EC 3.2.1.91)活性和/或木葡聚糖-特异性内切-β-1,4-葡聚糖酶(EC 3.2.1.151)活性和/或内切-β-1,6-半乳聚糖酶(EC 3.2.1.164)活性和/或β-1,3-甘露聚糖酶(EC 3.2.1.-)活性和/或阿拉伯糖基木聚糖-特异性内切-β-1,4-木聚糖酶(EC 3.2.1.-)活性和/或α-L-阿拉伯呋喃糖苷酶(EC 3.2.1.55)活性和/或阿魏酸酯酶(EC3.2.1.73)活性和/或α-葡糖醛酸酶(EC 3.2.1.115)活性等。
上述制备方法中,所述用所述复合酶制剂处理剩余的第一固体沉淀的方法为将剩余的第一固体沉淀加入50mM pH为2.0~9.0的缓冲液(所述缓冲液可以为柠檬酸、醋酸、磷酸、甘氨酸-盐酸缓冲液)进行重悬,再加入复合酶制剂获得酶解体系,加入比例为1%~5%(v/v,即所述复合酶制剂占酶解体系的体积百分比为1%~5%),在40℃~70℃条件下保温酶解处理24h。分别测定复合酶制剂水解葡聚糖底物、木聚糖底物的最适PH值曲线、最适温度曲线和温度稳定性曲线,得到复合酶制剂的催化条件为:水解木聚糖底物的最适pH为5.0,水解葡聚糖底物最适pH为5.0;水解木聚糖最适温度为55℃,水解葡聚糖最适温度为50℃,另外,在酶的温度稳定性实验中发现40-50℃时,酶活性比较稳定,55℃虽然活性高,但是半衰期短,酶活损失快。因此,用所述复合酶制剂处理剩余的第一固体沉淀的方法优选为将剩余的第一固体沉淀加入50mM pH为5.0的缓冲液进行重悬获得酶解体系,加入复合酶制剂,加入比例为1%~5%(v/v,即所述复合酶制剂占酶解体系的体积百分比为1%~5%),在40-50℃条件下保温酶解处理24h。
上述制备方法中,所述干燥为真空冷冻干燥。
上述制备方法中,还包括利用米糠制备米糠纤维的步骤。
上述制备方法中,所述第一固体沉淀的一部分与剩余的第一固体沉淀的质量比为1~2:3,优选的为1:3。
上述制备方法制备得到的米糠多糖也在本发明的保护范围之内。
本发明进一步提供了上述米糠多糖在提高乳化稳定性中的应用或在制备调味酱中的应用。
本发明进一步还提供了一种乳化稳定剂,所述乳化稳定剂的活性成分为上述米糠多糖。
本发明的有益效果如下:
本发明采用热水/超声浸提-复合酶-热水/超声浸提联用的方法对米糠纤维进行处理,通过热水浸提和超声浸提分两次破除植物细胞壁,使米糠多糖有效溶出,有助于后续米糠多糖的提取;再以热水和超声不能裂解的不溶性纤维为碳源,发酵培养具有完整纤维素、半纤维素降解酶系的裂褶菌,制备适合降解米糠纤维的复合酶制剂,并应用复合酶制剂处理米糠纤维;再次经过热水浸提和超声浸提不溶性米糠纤维,提高了米糠多糖的提取率和纯度。本发明米糠多糖的提取方法没有使用酸或碱液进行提取,绿色无污染,符合可持续发展的理念,适用范围广,能够大规模工业化生产。
附图说明
下面结合附图对本发明的具体实施方式作进一步详细的说明。
图1示出以米糠纤维为原料制备米糠多糖的步骤。
图2示出不同乳化稳定剂黏度的测定结果。
具体实施方式
为了更清楚地说明本发明,下面结合优选实施例和附图对本发明做进一步的说明。附图中相似的部件以相同的附图标记进行表示。本领域技术人员应当理解,下面所具体描述的内容是说明性的而非限制性的,不应以此限制本发明的保护范围。
实施例1以米糠纤维为原料制备米糠多糖
一种米糠多糖的制备方法,示意图如图1所示,具体步骤如下:
(1)米糠纤维的获得
取脱脂米糠,按固液比(脱脂米糠与水的质量比)为1:15的加入蒸馏水,调节pH值为6.0,向体系中加入耐高温α-淀粉酶,95℃水浴下处理30min,降低水浴温度为55℃,加入糖化酶,反应30min,并利用I2-KI溶液检查淀粉水解情况。向上述处理处理液中加入中性蛋白酶,在水浴温度55℃,pH为7.0的条件下,反应60min后,加热至100℃灭酶5min,纱布过滤,滤渣用蒸馏水洗涤2遍,105℃烘干获得米糠纤维,该米糠纤维为不溶性米糠纤维。
(2)热水浸提和超声浸提
将步骤(1)得到的米糠纤维浸于水中,料液比(米糠纤维与水的质量比)为1:15,进行热水浸提:在pH值调至3.0,温度90℃条件下浸提120min;再进行超声浸提:在提取温度70℃,超声功率为300W条件下浸提90min,得到提取液;将所述提取液固液分离,获得第一固体沉淀和第一上清液,对第一上清液进行真空冷冻干燥,得第一米糠多糖。
将第一固体沉淀按照质量比1:3分成两部分,即1/4第一固体沉淀和3/4第一固体沉淀。
(3)复合酶制剂制备
将1/4第一固体沉淀烘干、粉碎,作为碳源配制发酵培养基(所述碳源占所述发酵培养基的质量体积百分比为1%,w/v)发酵培养裂褶菌(Schizophyllum commune,购自中国普通微生物菌种保藏管理中心CGMCC 5.0819),培养条件:在温度30℃,转速220rpm的培养条件下发酵培养3天后,得到发酵液进行固液分离,用截留分子量5000Da中空纤维柱浓缩发酵液,浓缩10倍,制备成复合酶制剂;
其中,所述发酵培养基中除碳源外还加入了无机盐和余量的水,所述无机盐的组分及其占发酵培养基中的质量体积百分比为0.02%尿素,0.42%(NH4)2SO4,0.2%KH2PO4,0.03%CaCl2,0.03%MgSO4·H2O,0.2%吐温80和体积百分比为0.2%微量元素溶液,其中,所述微量元素溶液的组分及其占微量元素溶液中的质量体积百分比为0.5%FeSO4·7H2O,0.16%MnSO4·4H2O,0.14%ZnSO4·7H2O,0.2%CoCl2和余量的水。
发酵培养的时间是根据发酵过程中取样测定发酵液酶活(包括葡聚糖酶、木聚糖酶、α-L-阿拉伯呋喃糖苷酶、阿魏酸酯酶、α-葡糖醛酸酶)和蛋白含量确定的,当在温度30℃,转速220rpm的培养条件下发酵培养3天后发酵液酶活和蛋白含量最优。
分别测定复合酶制剂水解葡聚糖底物、木聚糖底物的最适pH值、最适温度和温度稳定性;最终得到复合酶制剂的催化条件为:水解木聚糖底物最适pH为5.0,水解葡聚糖底物最适pH为5.0;水解木聚糖最适温度为55℃,水解葡聚糖最适温度为50℃。
用iTRAQ蛋白质组分析技术测定浓缩发酵液中纤维降解酶的组成和含量,包含40个家族(GH1、GH2、GH3、GH5、GH6、GH7、GH10、GH11、GH13、GH15、GH16、GH17、GH18、GH25、GH27、GH28、GH30、GH31、GH35、GH37、GH43、GH45、GH47、GH51、GH53、GH55、GH61、GH62、GH71、GH74、GH76、GH79、GH81、GH88、GH92、GH93、GH95、GH105、GH115、GH125)的75个糖苷水解酶蛋白;具体的该复合酶制剂包含以下活性:内切-β-1,4-葡聚糖酶/纤维素酶(EC3.2.1.4)活性、内切-β-1,4-木聚糖酶(EC3.2.1.8)活性、β-葡糖苷酶(EC 3.2.1.21)活性、β-甘露糖苷酶(EC3.2.1.25)活性、葡聚糖β-1,3-葡糖苷酶(EC 3.2.1.58)活性、外切-β-1,4-葡聚糖酶/纤维糊精酶(EC 3.2.1.74)活性、葡聚糖内切-1,6-β-葡糖苷酶(EC 3.2.1.75)活性、甘露聚糖内切-β-1,4-甘露糖苷酶(EC 3.2.1.78)活性、纤维素β-1,4-纤维二糖苷酶(EC3.2.1.91)活性、木葡聚糖-特异性内切-β-1,4-葡聚糖酶(EC 3.2.1.151)活性、内切-β-1,6-半乳聚糖酶(EC 3.2.1.164)活性、β-1,3-甘露聚糖酶(EC 3.2.1.-)活性、阿拉伯糖基木聚糖-特异性内切-β-1,4-木聚糖酶(EC 3.2.1.-)活性、α-L-阿拉伯呋喃糖苷酶(EC 3.2.1.55)活性、阿魏酸酯酶(EC 3.2.1.73)活性和α-葡糖醛酸酶(EC3.2.1.115)活性。
(4)可溶性米糠多糖提取
将3/4第一固体沉淀加入pH为5.0缓冲液进行重悬,再加入复合酶制剂获得酶解体系(复合酶制剂/酶解体系的体积比为1/20(v/v)),在40-50℃条件下保温酶解处理24h;酶解处理后再次进行热水浸提和超声浸提,热水浸提是在pH值调至3.0,温度90℃条件下浸提120min,超声浸提是在提取温度70℃,超声功率300W的条件下提取90min,不断搅拌,固液分离获得第二上清液,对第二上清液进行真空冷冻干燥,得第二米糠多糖;
将第一米糠多糖和第二米糠多糖合并得到米糠多糖,得率12.49%,纯度98.1%。
也可以将第一上清液和第二上清液合并,经真空冷冻干燥得到米糠多糖,同样的,得率12.49%,纯度98.1%。
对比例1以脱脂米糠为原料制备米糠多糖
一种米糠多糖的制备方法,具体步骤如下:
(1)热水浸提和超声浸提
将脱脂米糠浸于水中,料液比为1:15,进行热水浸提:在pH值调至3.0、温度90℃条件下浸提120min;再进行超声浸提:在提取温度70℃,超声功率300W条件下提取90min得到提取液;将所述提取液固液分离,获得第一固体沉淀和第一上清液,对第一上清液进行真空冷冻干燥,得米糠多糖。
将第一固体沉淀按照质量比1:3分成两部分,即1/4第一固体沉淀和3/4第一固体沉淀。
(2)复合酶制剂制备
将1/4第一固体沉淀烘干、粉碎,作为碳源配制发酵培养基(所述碳源占所述发酵培养基的质量体积百分比为1%,w/v)发酵培养裂褶菌(Schizophyllumcommune,购自中国普通微生物菌种保藏管理中心CGMCC 5.0819),培养条件:在温度30℃,转速220rpm的培养条件下发酵培养3天后,得到发酵液进行固液分离,用截留分子量5000Da中空纤维柱浓缩发酵液,浓缩10倍,制备成复合酶制剂;
其中,所述发酵培养基中除碳源外还加入了无机盐和余量的水,所述无机盐的组分及其占发酵培养基中的质量体积百分比为0.02%尿素,0.42%(NH4)2SO4,0.2%KH2PO4,0.03%CaCl2,0.03%MgSO4·H2O,0.2%吐温80和体积百分比为0.2%微量元素溶液,其中,所述微量元素溶液的组分及其占微量元素溶液中的质量体积百分比为0.5%FeSO4·7H2O,0.16%MnSO4·4H2O,0.14%ZnSO4·7H2O,0.2%CoCl2和余量的水。
(3)可溶性米糠多糖提取
将3/4第一固体沉淀加入pH为5.0缓冲液进行重悬,加入裂褶菌发酵浓缩复合酶,按照1/20(v/v,复合酶/米糠提取液体积)比例添加,在40-50℃条件下保温酶解处理24h;酶解处理后再次进行热水浸提和超声浸提,热水浸提是在pH值调至3.0,温度90℃条件下浸提120min,超声浸提是在提取温度70℃,超声功率为300W,提取时间90min,不断搅拌,固液分离获得第二上清液,对第二上清液进行真空冷冻干燥,得米糠多糖。
将第一上清液和第二上清液合并,经真空冷冻干燥,得米糠多糖,得率28.6%,纯度63.64%。
对比例2未使用复合酶制备米糠多糖
将实施例1步骤(1)获得的米糠纤维浸于水中,料液比为1:15,进行热水浸提,即将米糠纤维,pH值调至3.0,在90℃条件下浸提120min;再进行超声浸提:在提取温度70℃,超声功率300W条件下提取90min得到提取液;将所述提取液固液分离,获得上清液进行真空冷冻干燥,得米糠多糖,得率7.88%,纯度93.5%。
试验例 乳化稳定性的测定
将实施例1中米糠纤维提取得到的米糠多糖与对比例1中米糠提取得到的米糠多糖、果胶、醋酸酯变性淀粉四种乳化稳定剂进行单体之间乳化稳定性的对比实验。
分别称取每种乳化稳定剂样品溶解于盐:色拉油:水=15:20:65的调味酱模拟体系的溶液中,配制成相应质量分数的乳液(2%对比例1中米糠提取得到的米糠多糖、2%果胶、2%醋酸酯变性淀粉、2%实施例1中米糠纤维提取得到的米糠多糖),沸水浴中糊化并保温15min,剪切分散乳化3min,然后自然冷却至室温,分别测定黏度、凝沉性、乳化能力。
具体测定方法和结果如下:
用旋转流变仪(AR2000ex型)测定其黏度。选用直径为40mm的平行板测量系统,将样品台温度设定为30℃,板间距1.0mm,将剪切速率从0.1s-1增加到600s-1,结果见图1:2%对比例1中米糠提取得到的米糠多糖(图中以“2%米糠提取多糖”表示)与2%果胶、2%醋酸酯变性淀粉表现出相似的黏度;2%实施例1中米糠纤维提取得到的米糠多糖(图中以“2%米糠纤维水解多糖”表示)表现出较低的黏度。
移取乳液50ml离心管中,以3000rpm离心15min,记录沉淀层重量。凝沉性(%)=沉淀层体积/离心管糊液的总体积×100%,凝沉性越接近0(N/A),样品的乳化稳定性越好。乳化能力(%)=离心管中表层溶液重量/离心管中液体总重×100%。凝沉性与乳化能力的结果如表1所示,2%实施例1中米糠纤维提取得到的米糠多糖(图中以“2%米糠纤维水解多糖”表示)的乳化能力明显优于2%果胶、2%醋酸酯变性淀粉、2%对比例1米糠提取得到的米糠多糖(表中以“2%米糠提取多糖”表示)。
表1不同稳定剂的凝沉性和乳化能力
项目 2%米糠纤维水解多糖 2%米糠提取多糖 2%果胶 2%醋酸酯变性淀粉
凝沉性/(%) N/A N/A N/A N/A
乳化能力/(%) 32.06 20.5 3.6 5.5
综上所述,本发明实施例1中米糠纤维提取得到的米糠多糖相对于果胶、醋酸酯变性淀粉、对比例1米糠提取得到的米糠多糖具有较强的乳化稳定性。
显然,本发明的上述实施例仅仅是为清楚地说明本发明所作的举例,而并非是对本发明的实施方式的限定,对于所属领域的普通技术人员来说,在上述说明的基础上还可以做出其它不同形式的变化或变动,这里无法对所有的实施方式予以穷举,凡是属于本发明的技术方案所引伸出的显而易见的变化或变动仍处于本发明的保护范围之列。

Claims (13)

1.一种米糠多糖的制备方法,其特征在于:所述制备方法包括如下步骤:
将米糠纤维浸于水进行热水浸提和超声浸提,获得第一提取液;
将所述第一提取液进行固液分离,获得第一固体沉淀和第一上清液,对第一上清液进行干燥,得第一米糠多糖;
将第一固体沉淀的一部分进行烘干和粉碎后作为发酵培养基的碳源发酵培养裂褶菌,获得发酵液,将所述发酵液进行固液分离后浓缩成复合酶制剂;
用所述复合酶制剂处理剩余的第一固体沉淀,再次进行所述热水浸提和超声浸提,获得第二提取液;
将所述第二提取液进行固液分离,获得第二上清液,对第二上清液进行干燥,得第二米糠多糖;
将第一米糠多糖和第二米糠多糖合并得到米糠多糖。
2.一种米糠多糖的制备方法,其特征在于:所述制备方法包括如下步骤:
将米糠纤维浸于水进行热水浸提和超声浸提,获得第一提取液;
将所述第一提取液进行固液分离,获得第一固体沉淀和第一上清液;
将第一固体沉淀的一部分进行烘干和粉碎后作为发酵培养基的碳源发酵培养裂褶菌,获得发酵液,将所述发酵液进行固液分离后浓缩成复合酶制剂;
用所述复合酶制剂处理剩余的第一固体沉淀,再次进行所述热水浸提和超声浸提,获得第二提取液;
将所述第二提取液进行固液分离,获得第二上清液;
将所述第一上清液和第二上清液合并后进行干燥得到米糠多糖。
3.根据权利要求1或2所述的制备方法,其特征在于:所述热水浸提为在pH 3.0、温度为80℃~100℃的条件下浸提120min。
4.根据权利要求3所述的制备方法,其特征在于:所述米糠纤维与水的质量比为1:8~1:15。
5.根据权利要求1或2所述的制备方法,其特征在于:所述超声浸提为在温度为70℃~100℃、超声功率为150W~300W的条件下浸提90min。
6.根据权利要求1或2所述的制备方法,其特征在于:所述发酵培养是在温度为30℃,转速为220rpm的培养条件下发酵培养2~10天。
7.根据权利要求6所述的制备方法,其特征在于:所述发酵培养基除了碳源之外还加入了无机盐和水,所述碳源的质量占所述发酵培养基的总体积的1~10%。
8.根据权利要求1或2所述的制备方法,其特征在于:所述浓缩为浓缩2~10倍。
9.根据权利要求1或2所述的制备方法,其特征在于:所述用所述复合酶制剂处理剩余的第一固体沉淀的方法为将剩余的第一固体沉淀加入50mM pH为2.0~9.0的缓冲液中进行重悬,再加入复合酶制剂获得酶解体系,在40℃~70℃条件下保温酶解处理24h。
10.根据权利要求9所述的制备方法,其特征在于:所述复合酶制剂占酶解体系的体积百分比为1%~5%。
11.权利要求1-10任一所述的制备方法制备得到的米糠多糖。
12.权利要求11所述的米糠多糖在如下任一中的应用:
1)在提高乳化稳定性中的应用;
2)在制备调味酱中的应用。
13.一种乳化稳定剂,其特征在于,所述乳化稳定剂的活性成分为权利要求11所述的米糠多糖。
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