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用一种宏观的体外方法降解两种可溶性蛋白——酪蛋白和卵蛋白。

Degradation of two soluble proteins--casein and egg protein by a macro in vitro method.

机构信息

Department of Animal Nutrition & Management, Kungsängen Research Centre, Swedish University of Agricultural Sciences, Uppsala, Sweden.

出版信息

J Anim Physiol Anim Nutr (Berl). 2013 Aug;97(4):656-65. doi: 10.1111/j.1439-0396.2012.01306.x. Epub 2012 Apr 28.

Abstract

Degradation of casein and egg protein was studied with whole rumen contents (RC) in a macro in vitro system to elucidate previous findings of initial rapid disappearance of soluble proteins in vitro. Five to 7.5 kg of RC from a dry and/or a lactating cow were incubated with buffer and casein or egg protein for 180 min with frequent sampling. Degradation was measured as loss of trichloroacetic acid precipitable N (TCA-N) from the inocula. Normal (39°C) and low (2°C) temperature incubations were examined in Exp. 1, using 1 g of TCA-N from casein. Four levels of casein (0-12 g TCA-N) in Exp. 2 and four levels of egg albumin (0-24 g TCA-N) in Exp. 3 were fermented at 39°C. Initial recovery of casein TCA-N was 106% at 2°C and 56% at 39°C (Exp. 1). Casein (TCA-N) recovered initially increased in Exp. 2 from 21% at 3 g to 86% at 12 g TCA-N, while absolute loss remained relatively constant at 358 mg TCA-N/kg RC (SD=47). Fractional degradation rate was highest (0.03/min) at the intermediate dosage level. In the absence of rumen fluid (Exp. 4), no casein was lost. Initial egg protein recovery was on average 103% (Exp. 3). Recovery seemed unaffected by dosage level, and absolute degradation rate was relatively constant over time and increased with dosage level (p<0.001) from 1.48 to 2.95 mg TCA-N/(kg RC × min). Maximum degradation rate [mg TCA-N/(kg RC × min)] and affinity constant (mg TCA-N/kg RC) were estimated at 261 and 1650, respectively. It is concluded that a surprisingly constant amount of casein disappears immediately from warm rumen fluid and that this does not occur either with chilled RC, in the absence of rumen fluid, or when replaced with egg protein. The mechanisms for this disappearance are yet to be discovered.

摘要

本研究采用全瘤胃液在宏观体外体系中研究了酪蛋白和卵蛋白的降解,以阐明体外可溶性蛋白最初快速消失的先前发现。从干奶牛和/或泌乳奶牛中采集 5 至 7.5kg 的瘤胃液,与缓冲液和酪蛋白或卵蛋白一起孵育 180 分钟,频繁取样。降解通过接种物中三氯乙酸沉淀氮(TCA-N)的损失来测量。在实验 1 中,使用 1g 酪蛋白 TCA-N 分别在正常(39°C)和低温(2°C)下孵育,在实验 2 中用 4 个酪蛋白水平(0-12gTCA-N)和实验 3 中用 4 个卵白蛋白水平(0-24gTCA-N)在 39°C 下发酵。在 2°C 时初始酪蛋白 TCA-N 的回收为 106%,在 39°C 时为 56%(实验 1)。实验 2 中,初始酪蛋白 TCA-N 的回收从 3g 时的 21%增加到 12gTCA-N 时的 86%,而绝对损失相对恒定,为 358mgTCA-N/kgRC(SD=47)。在中间剂量水平时,分级降解率最高(0.03/min)。在没有瘤胃液的情况下(实验 4),没有酪蛋白丢失。初始卵蛋白回收率平均为 103%(实验 3)。回收率似乎不受剂量水平的影响,绝对降解率在时间上相对恒定,并随剂量水平增加(p<0.001),从 1.48 增加到 2.95mgTCA-N/(kgRC×min)。最大降解率[mgTCA-N/(kgRC×min)]和亲和常数(mgTCA-N/kgRC)分别估计为 261 和 1650。综上所述,令人惊讶的是,相当数量的酪蛋白会立即从温暖的瘤胃液中消失,而这种情况不会发生在冷藏的 RC 中,也不会在没有瘤胃液的情况下发生,或者当用卵蛋白替代时也不会发生。这种消失的机制尚待发现。

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