Armstead I P, Ling J R
Department of Biochemistry, School of Life Sciences, University of Wales, United Kingdom.
Appl Environ Microbiol. 1993 Oct;59(10):3360-6. doi: 10.1128/aem.59.10.3360-3366.1993.
Mixed ruminal bacteria, isolated from sheep (Q and W) fed a concentrate and hay diet, were anaerobically incubated with either 14C-peptides or 14C-amino acids. Experiment 1 showed that uptake of both 14C-labeled substrates was rapid, but the rate for amino acids was twofold greater than for peptides (molecular weight, 1,000 to 200) initially but was similar after 10 min. Experiment 2 demonstrated that metabolism was also rapid; at least 90% of either 14C-labeled substrate was metabolized by 3 min. Of the radioactivity remaining in bacteria, approximately 30% was in the form of 14C-amino acids, but only in leucine, tyrosine, and phenylalanine. Supernatant radioactivity was contained only in tyrosine, phenylalanine, and mostly proline for incubations with 14C-amino acids but in up to 10 amino acids when 14C-peptides were the substrates. Short-term incubations (< 5 min; experiment 3) confirmed previous uptake patterns and showed that the experimental system was responsive to substrate competition. Experiment 4 demonstrated that bacteria from sheep Q possessed initial and maximum rates of 14C-amino acid uptake approximately fourfold greater (P < 0.01) than those of 14C-peptides, but with no significant differences (P > 0.1) between four 14C-peptide substrate groups with molecular weights of 2,000 to < 200. By contrast, bacteria from sheep W showed no such distinctions (P > 0.1) between rates for 14C-peptides and 14C-amino acids. Calculations suggested that peptides could supply from 11 to 35% and amino acids could supply from 36 to 68% of the N requirements of mixed ruminal bacteria.(ABSTRACT TRUNCATED AT 250 WORDS)
从喂食精饲料和干草日粮的绵羊(Q和W)中分离出的混合瘤胃细菌,与14C标记的肽或14C标记的氨基酸进行厌氧培养。实验1表明,两种14C标记底物的摄取都很快,但氨基酸的摄取速率最初是肽(分子量为1000至200)的两倍,但10分钟后相似。实验2表明,代谢也很快;3分钟内至少90%的14C标记底物被代谢。细菌中剩余的放射性中,约30%是以14C氨基酸的形式存在,但仅存在于亮氨酸、酪氨酸和苯丙氨酸中。与14C氨基酸孵育时,上清液放射性仅存在于酪氨酸、苯丙氨酸中,且大部分为脯氨酸;而以14C肽为底物时,上清液放射性存在于多达10种氨基酸中。短期孵育(<5分钟;实验3)证实了先前的摄取模式,并表明实验系统对底物竞争有反应。实验4表明,绵羊Q的细菌对14C氨基酸的初始摄取速率和最大摄取速率比14C肽大约高四倍(P<0.01),但分子量为2000至<200的四个14C肽底物组之间没有显著差异(P>0.1)。相比之下,绵羊W的细菌在14C肽和14C氨基酸的摄取速率之间没有这种差异(P>0.1)。计算表明,肽可为混合瘤胃细菌提供11%至35%的氮需求,氨基酸可为其提供36%至68%的氮需求。(摘要截短于250字)