物理預(yù)處理—可控酶解聯(lián)合改性蠶蛹蛋白的研究
發(fā)布時(shí)間:2019-06-18 17:22
【摘要】:我國(guó)蠶業(yè)歷史悠久,是世界上最大的蠶絲產(chǎn)地。由于傳統(tǒng)的栽桑養(yǎng)蠶以獲取蠶絲為主要目的,蠶蛹就成了主要副產(chǎn)品,我國(guó)每年可產(chǎn)新鮮蠶蛹50萬(wàn)噸以上,相當(dāng)長(zhǎng)的一段時(shí)期內(nèi)蠶蛹基本上都被用作飼料,導(dǎo)致了巨大資源浪費(fèi)。為了提高蠶蛹蛋白的利用價(jià)值,本研究采用微生物發(fā)酵法對(duì)蠶蛹的異味進(jìn)行脫除,之后利用超微粉碎處理-酶解和超聲波處理-酶解聯(lián)合改性的方法對(duì)蠶蛹蛋白進(jìn)行功能特性的改良,并研究改性前后的營(yíng)養(yǎng)價(jià)值和生物學(xué)活性,為高附加值利用蠶蛹提供理論依據(jù)。以發(fā)酵后蠶蛹的異味物質(zhì)為指標(biāo),從8種菌株中篩選出的白色假絲酵母的異味脫除效果最好;通過(guò)單因素試驗(yàn),篩選出了最佳接種量為2%,最佳發(fā)酵溫度為35℃,最佳發(fā)酵時(shí)間為40h;通過(guò)GC-MS對(duì)不同發(fā)酵工藝下蠶蛹的氣味物質(zhì)進(jìn)行了分析,結(jié)果顯示經(jīng)發(fā)酵后蠶蛹的異味成分基本檢測(cè)不到。經(jīng)過(guò)微細(xì)化處理、酶解處理和微細(xì)化-酶解處理后,蠶蛹蛋白的溶解度分別為未處理的1.84、2.32和2.69倍,乳化性分別提高3.6%、15.05%和21.52%,起泡性及起泡穩(wěn)定性也不同程度地得到提高。此外,經(jīng)微細(xì)化處理后蠶蛹蛋白的水解度可達(dá)到22.59%,為未處理的1.58倍,蛋白回收率可達(dá)到56.68%,為未處理的1.82倍。不同處理改性的蠶蛹蛋白的分子變小,二硫鍵含量最高可降低19.70%,巰基含量最高可升高55.30%。改性后蠶蛹蛋白二級(jí)結(jié)構(gòu)發(fā)生巨大變化,其中α-螺旋、β-折疊和β-轉(zhuǎn)角等結(jié)構(gòu)極度減少,無(wú)規(guī)則卷曲結(jié)構(gòu)大量增加,最終蛋白的二級(jí)結(jié)構(gòu)以無(wú)規(guī)則卷曲為主。經(jīng)過(guò)超聲波處理、酶解處理和超聲波-酶解處理后,蠶蛹蛋白的溶解度分別為未處理的2.03、2.32和3.59倍,乳化性分別提高10.94%、15.05%和40.4%,起泡性及起泡穩(wěn)定性也不同程度地得到提高。此外,經(jīng)超聲波處理后蠶蛹蛋白的水解度可達(dá)到24.55%,為未處理的1.72倍,蛋白回收率可達(dá)到75.99%,為未處理的2.44倍。不同處理改性的蠶蛹蛋白的分子變小,二硫鍵含量最高可降低20.39%,巰基含量最高可升高59.59%。改性后蠶蛹蛋白二級(jí)結(jié)構(gòu)發(fā)生巨大變化,其中α-螺旋、β-折疊和β-轉(zhuǎn)角等結(jié)構(gòu)極度減少,無(wú)規(guī)則卷曲結(jié)構(gòu)大量增加,最終蛋白的二級(jí)結(jié)構(gòu)主要是無(wú)規(guī)則卷曲。改性后蠶蛹蛋白的氨基酸分析、營(yíng)養(yǎng)學(xué)評(píng)價(jià),抗氧化活性、體外血管緊張素轉(zhuǎn)化酶(ACE)抑制率和促小鼠脾細(xì)胞增殖能力等研究結(jié)果表明,改性后蠶蛹蛋白的必需氨基酸(EAA)與非必需氨基酸(NEAA)的比值(EAA/NEAA)相比未處理蠶蛹蛋白提高了10%以上,均達(dá)到了理想蛋白模式,最高值達(dá)到了66.86%;必需氨基酸/總氨基酸的比值也均高于聯(lián)合國(guó)糧食及農(nóng)業(yè)組織/世界衛(wèi)生組織(FAO/WHO)推薦模式的36%,氨基酸評(píng)分(AAS)及必需氨基酸指數(shù)(EAAI)也都得到了一定程度的提高,EAAI值均高于90%。改性后蠶蛹蛋白的1,1-二苯基-2-三硝基苯肼(DPPH)清除率最高達(dá)70%,亞鐵離子的螯合率最高達(dá)到80%,總還原力也有較大的提升,相比未處理蠶蛹蛋白,改性后蠶蛹蛋白的抗氧化能力得到顯著提升。改性后蠶蛹蛋白對(duì)血管緊張素轉(zhuǎn)化酶(ACE)抑制率的IC50最低為2.00mg/mL,僅為未處理的5.93%。在劑量為50μg/mL時(shí),改性后蠶蛹蛋白促小鼠脾細(xì)胞增殖能力可增加100%。說(shuō)明改性后蠶蛹蛋白的營(yíng)養(yǎng)價(jià)值、抗氧化活性、體外ACE抑制率以及促小鼠脾細(xì)胞增殖能力都得到了較為明顯地提高。
[Abstract]:China's silkworm industry has a long history and is the world's largest silk production area. Due to the traditional mulberry-growing silkworm to obtain silk as the main purpose, the silkworm chrysalis is the main by-product, and the silkworm chrysalis can produce more than 500,000 tons of fresh silkworm chrysalis per year, and the silkworm chrysalis is basically used as a feed for a long period of time, leading to a great resource waste. in order to improve the utilization value of the silkworm chrysalis protein, the research adopts a microbial fermentation method to remove the peculiar smell of the silkworm chrysalis, And the nutritional value and the biological activity before and after modification are studied, and the theoretical basis is provided for the use of the silkworm chrysalis for high added value. The peculiar smell removal effect of Candida albicans from 8 strains was the best, and the optimal fermentation temperature was 35 鈩,
本文編號(hào):2501657
[Abstract]:China's silkworm industry has a long history and is the world's largest silk production area. Due to the traditional mulberry-growing silkworm to obtain silk as the main purpose, the silkworm chrysalis is the main by-product, and the silkworm chrysalis can produce more than 500,000 tons of fresh silkworm chrysalis per year, and the silkworm chrysalis is basically used as a feed for a long period of time, leading to a great resource waste. in order to improve the utilization value of the silkworm chrysalis protein, the research adopts a microbial fermentation method to remove the peculiar smell of the silkworm chrysalis, And the nutritional value and the biological activity before and after modification are studied, and the theoretical basis is provided for the use of the silkworm chrysalis for high added value. The peculiar smell removal effect of Candida albicans from 8 strains was the best, and the optimal fermentation temperature was 35 鈩,
本文編號(hào):2501657
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