高活性納豆激酶工藝改進(jìn)研究

張浩 王家林

摘要[目的]優(yōu)化高活性納豆激酶的納豆生產(chǎn)工藝條件。[方法]從市售納豆產(chǎn)品中分離篩選出高活性的菌株作為試驗(yàn)菌株，采用固態(tài)發(fā)酵的方法制備納豆，對(duì)影響納豆激酶產(chǎn)量的主要因素浸泡時(shí)間、接種量、發(fā)酵時(shí)間等進(jìn)行考察，研究高活性納豆激酶的工藝條件。[結(jié)果]利用對(duì)甲基苯磺酰L精氨酸甲酯（TAME）法測(cè)定納豆激酶活性，確定最佳納豆激酶酶活的納豆生產(chǎn)工藝條件為浸泡時(shí)間8 h、接種量10%、發(fā)酵時(shí)間48 h。[結(jié)論]該研究可為高活性納豆激酶的生產(chǎn)提供參考。

關(guān)鍵詞納豆；納豆激酶；活性測(cè)定；工藝優(yōu)化

中圖分類號(hào)TQ925文獻(xiàn)標(biāo)識(shí)碼

A文章編號(hào)0517-6611（2017）10-0093-02

Study on the Improvement of High Activity Natto Kinase

ZHANG Hao，WANG Jialin（Biological Engineering Lab of Qingdao University of Science and Technology，Qingdao，Shandong 266042）

Abstract[Objective] To optimize the process conditions of high activity natto kinase.[Method]Natto was isolated from commercial natto products and used as the experimental strain.Natto was prepared by solidstate fermentation.The process conditions of high activity natto kinase were studied.The main factors affecting the yield of natto kinase were as follows：immersion time，inoculation amount and fermentation time.[Result]The activity of natto kinase was determined by TAME method.The optimum natto kinase production process conditions were determined as 8 h soaking time，10% inoculation amount，48 h fermentation time.[Conclusion] The study can provide reference for production of high activity natto kinase.

Key wordsNatto；Natto kinase；Activity determination；Process optimization

納豆是經(jīng)過蒸煮的大豆在納豆芽孢桿菌的發(fā)酵作用下產(chǎn)生的發(fā)酵食品[1]。納豆含有多種生理活性物質(zhì)，具有擴(kuò)張血管、溶解血栓、促進(jìn)血液循環(huán)、抗癌、抗菌、抗氧化、延緩衰老、預(yù)防骨質(zhì)疏松、改善腸道環(huán)境等生理功能。納豆激酶是由納豆芽孢桿菌分泌的具有溶栓作用的堿性絲氨酸蛋白酶[2]，是由275個(gè)氨基酸殘基組成的單鏈多肽，屬于胞外酶，其相對(duì)分子質(zhì)量不大，具有溶解血栓的作用，并且在人體內(nèi)作用迅速、半衰期短，因此近年來受到世界各國(guó)醫(yī)學(xué)界的廣泛關(guān)注。

該試驗(yàn)是在成品納豆中提取納豆芽孢桿菌，經(jīng)過逐級(jí)放大培養(yǎng)后接種到高溫蒸汽滅菌的黃豆中，通過改變浸泡時(shí)間、接種量、發(fā)酵時(shí)間、后熟時(shí)間等條件進(jìn)行發(fā)酵培養(yǎng)。將納豆碾碎后加生理鹽水溶解，通過冷凍離心保留上清液，再經(jīng)過2次硫酸銨沉淀分離提純得到納豆激酶粗酶，再利用pH 6.5的磷酸緩沖液溶解得到粗酶液[3]。……