kg, H 2O 14 g, KH 2PO 4 2 g,n H 4C l 3 g, KC l 5 g, CaC l2
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- Ωρίων Μπουκουβαλαίοι
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1 34 7 () V o l. 34 N o Jour. of N o rthw est Sci2T ech U niv. of A gri. and Fo r. (N aṫ Sci. Ed. ) July 2006 Ξ 1a, 2, 1a, 1b, 1b (1 a., b., ; 2, ) [ ], L 2 (M et) ( ) L 2 ( ), 2 (A ST ) (AL T ), M et A ST AL T, M et (Gly) (M et), (Cys2s) (Ser) (P < 0. 05), 16 h (H is) 8 h (P < 0. 05), (V al),,v al, Cys2s, Ser H is M et [ ] ; ; (A ST ); (AL T ); [] S [] A [ ] (2006) ,,,, Zikak is [1 ] W ak ita [2 ] ; O r2 rash id [3 ],,, ; Cu rtis [4 ],,,,,,,, [5 ] ; [2 ] ; Ca + + (GO T ) (GPT ) [6 ], L 2 (M et), (A ST ) (AL T ), : 800 ggkg, 120 ggkg, 63 ggkg, 10 ggkg, 3 ggkg, 4 ggkg 3, 2 (09: 00 17: 00), kg, kg, 6,, 4, L ee [7 ], 1. 2 Su sane [8 ] :M gc l2 6H 2O 14 g, KH 2PO 4 2 g,n H 4C l 3 g, KC l 5 g, CaC l2 Ξ [ ] [ ] ( ) [ ] (1956- ),,,,, E2m ail: lilum u@m ail. hf. ah. cn [ ] (1956- ),,,,,, E2m ail: sytu. edu. cn
2 7 : 23 2H 2O 0. 6 g, ml g 10 ml, mo lgl N aoh ml mmo lgl H EPES 1, g, 0. 2 g, 0. 2 g, B g, B g, B g, g, g, B g, g Λm mo lgl HC l M nc l2 4H 2O g, N ic l2 6H 2O g, N am oo 2 6H 2O g, H 3BO g, FeSO 4 7H 2O g, CoC l2 6H 2O g, SeO g, N avo 2 6H 2O g, CuC l2 2H 2O g ml, 10 ml, 10 ml, 1 ggl 1 ml, K 2H PO g, 830 ml, 1 mo lgl KOH ph ml, N a2co 3 4 g, ml,, ( 95% N 2+ 5% CO 2) 5 m in, N a2s 9H 2O L g, ( ), 100 ml,,, 20 m in1 ml , 5 ml, mo lgl L 2 (M et, ) 1 ml,, 3 L 2, , 8 16 h, 2 ml, 2, 1 4% ( rgm in, 30 m in), - 20 ; 1 A ST AL T 1100, A ST AL T , 1. 5 (x s), SA S (9. 0 ) [9 ], M et A ST AL T L 2 ( ) (A ST, AL T ) 1 1, 0 h, 8 h,, 16 h (T rp ) (Phe) 2, T rp 0, 8 h (P < 0. 01), Phe (1. 04 m ggkg) 0 h (4. 58 m ggkg) (P < 0. 05) T rp, Phe,, Phe, Phe, Phe [10211 ], Phe - (GPA ase), Phe GPA ase 1, M et 8 h, A ST 0 h, 16 h 5. 0 Λmo lg(l m in - 1 ), 0 16 h A ST AL T 0 h, 6. 7 Λmo lg(l m in - 1 ),, 8 h 5. 3 Λmo lg(l m in - 1 ), 0 h (P < 0. 05), 16 h 5. 0 Λmo lg(l m in - 1 ), 0 h (P < 0. 01), 8 h 16 h AL T,, A ST AL T, 2, 8 h 16 h (P < 0. 05), Glu A la
3 24 () 34 1 L 2 (M et) A ST AL T T able 1 Change of free am ino acid demonstration,a ST and AL T of culture fluid w ith no L 2m eth ionine added Item gh Incubation tim e A STg[Λmo l (L m in - 1 ) ] a b ab AL T g[λmo l (L m in - 1 ) ] A a b Bb L 2 g(m g kg - 1 ) M et g(m g kg - 1 ) L ys g(m g kg - 1 ) Cys2s g(m g kg - 1 ) Glu g(m g kg - 1 ) Ser g(m g kg - 1 ) H is g(m g kg - 1 ) Gly g(m g kg - 1 ) T h r g(m g kg - 1 ) A rg g(m g kg - 1 ) A la g(m g kg - 1 ) T yr g(m g kg - 1 ) T rp A A B g(m g kg - 1 ) Phe a ab b g(m g kg - 1 ) Ile g(m g kg - 1 ) L eu g(m g kg - 1 ) P ro g(m g kg - 1 ) V al g(m g kg - 1 ) To tal am ino acid : (P < 0. 05), (P < 0. 01) N o te: V alues w ithin a row w ith different sm all superscrip ts, different cap ital superscrip ts rep resent significant difference (P < 0. 05) and highly significant difference (P < 0. 01) respectively. The sam e is below M et A ST AL T L 2 (M et), A ST AL T M et A ST AL T T able 2 Change of free am ino acid demonstration,a ST and AL T of culture fluid w ith L 2m eth ionine added Item gh Incubation tim e A STg[Λmo l (L m in - 1 ) ] a ab b AL T g[λmo l (L m in - 1 ) ] a b b L 2 g(m g kg - 1 ) M et g(m g kg - 1 ) L ys g(m g kg - 1 ) Cys2s a ab b g(m g kg - 1 ) Glu g(m g kg - 1 ) Ser a ab b g(m g kg - 1 ) H is ab a b g(m g kg - 1 ) Gly g(m g kg - 1 ) T h r g(m g kg - 1 ) A rg g(m g kg - 1 ) A la g(m g kg - 1 ) T yr g(m g kg - 1 ) T rp g(m g kg - 1 ) Phe g(m g kg - 1 ) Ile g(m g kg - 1 ) L eu g(m g kg - 1 ) P ro g(m g kg - 1 ) V al g(m g kg - 1 ) To tal am ino acid
4 7 : 25 2, (Cys2s) (Ser) (H is) 0 h, (Cys2 s) m ggkg, Cys2s, 16 h m ggkg, 0 h (P < 0. 05) (Ser) Cys2s M et 2, [12 ] 2, 8 h, (H is) 0 h m ggkg m ggkg,, 16 h, m ggkg, 8 h (P < 0. 05) H is,, M et V al, V al, M et T rp Phe, M et 2, M et A ST, 16 h A ST 0 h AL T, 8 16 h AL T 0 h 2. 3 M et A ST, AL T, M et, M et A ST AL T M et, M et M et, M et, Gly,M et V al, M et M et, Gly M et,v al M et, M et, V al Gly, A ST AL T, 3, M et V al, Ser, Cys2s H is, H is, Gly, A ST AL T V al, Ser, Cys2s H is M et, M et, M et [ ] [ 1 ] Zikak is P J, Salsbury R L. M etabo lism of sulfur am ino by rum en[j ]. J D airy Sci, 1969, 52 (12) : [2 ] W ak ita M, Ho sh ino S. A branched2chain am ino acid am ino transferase from the rum en ciliate genus entodinium [J ]. J P ro tozoo l, 1975, 22 (2) : [ 3 ] O r2rash id M M, O nodera R,W adud S. Studies on the utilization sulfoxide and m eth ionine sulfone by rum en m icroo rganism s in v itra [J ]. Am ino A cid, 2003, 24: [ 4 ] Curtis S, Russell N, Chalupa W. D egradation of am ino acids by pure cultures of rum en bacteria[j ]. J A nim Sci, 1976, 43 (4) : [ 5 ] M o loney A P, Read M P, Keane M G. Effects of ardacin supp lem entation on rum en ferm entation and p ro tein degradability in steers[j ]. A nim al Feed Science and T echno logy, 1996, 57: [ 6 ] Faixova Z, Faix S. Infuence of m etal ions on rum inal enzym e activities[j ]. A cta V etbrno, 2002, 71: [ 7 ] L ee S S, H a J K, Cheng K J. Relative contribution of bacteria, p ro tozoa, and fungi to in v itro degradation of o rchard grass cell w all and their interactions[j ]. A pp lied and Environm entalm icrobio logy, 2000, 66 (9) : [ 8 ] Susane E L,M ichael K T, A nthony P J T, et al. Grow th of anaerobic rum en fungi on defined and sem i2defined m edia lack ing rum en fluid [J ]. J GeneralM icrobio logy, 1985, 131: [ 9 ] SA S Institute Inc. SA S U ser Guide: Statistics[M ]. V ersion 6, 4th Edition. N o rth Caro lina: SA S Institute Inc C ry, [ 10 ] Am in M R, O nodera R. Synthesis of phenylalanine and p roduction of o ther related compounds from phenylpyruvic acid and phenylacetic by rum inal bacteria, p ro tozoa and their m ixture in v itro[j ]. J Gen A pp lm icrobio l, 1997, 43: [ 11 ] Am in M R, O nodera R, Khan R I, et al. Purification and p roperties of glutanm ate2phenylpyruvate am ino transferase from rum en bacteri2 um p revo tella albensis[j ]. P sk istan J B io l Sci, 2001, 4 (11) :
5 26 () 34 [ 12 ],,,. : [M ]. :, Effect of m eth ion ine as N and C sou rces on am ino acids m etabo lism of rum en m icroo rgan ism in v itro L IL -m u 1a, 2,L E Guo-we i 1a, 1b, SH IY ong-hui 1b (1 a. K ey L aboratory of Education M inistry of Food S cience and S af ety; b. S chool of Food S cience and T echnology, S ou thern Y ang tz e U niversity,w ux i, J iang su , Ch ina; 2 S chool of A nim al S cience and T echnology,a nhu i A g ricu ltu ral U niversity, H ef ei,a nhu i , Ch ina) Abstract: In v itro studies w ere conducted to exam ine the effect of m eth ion ine u sed as so le n itrogen and carbon sou rces on the aspartate am ino tran sferase (A ST ), alan ine am ino tran sferase (AL T ) and am ino acids m etabo lism of rum en m icroo rgan ism s. R um en m icroo rgan ism s w ere co llected from fistu lated goats fed w ith alfalfa hay and a concen trate m ix tu re tw ice a day. Six serum bo ttles, each con tain ing 100 ml m ineral and vitam in m edia, taken and in jected in to the rum en flu id (off2feed granu les) 5 ml from every bo ttle, w ere random ly divided in to tw o group s,w ith th ree bo ttles in each group. O ne group w as in jected 0. 1 mmo lgl L 2 M et as trial group 1; the o ther th ree bo ttles w ere in jected w ithou t L 2M et, as a con tro l group. T hen they w ere anaerob ically incubated at 39 fo r 16 h. F ree am ino acids and tran sam inase (A ST,AL T ) in the su2 pernatan ts of the incubation w ere analyzed by H PL C and an au tom atic b iochem istry analyzer respectively. W hen M et w as u sed as a un ique sou rce of N and C of rum en m icroo rgan ism s, the con ten ts of free valine (V al) and h istidine (H is) w ere h igh ly sign ifican tly (P < 0. 01) and sign ifican tly (P < 0. 05) reduced respec2 tively; bu t glycine (Gly) w as h igh ly increased (P < 0. 01). N o sign ifican t differences in the o ther free am ino acids w ere found, as w ell as fo r GO T and GPT activities. T he resu lts indicate that V al and cysteine (Cys) are p robab ly the mo st impo rtan t essen tial am ino acids of rum en m icroo rgan ism s w hen M et serves as a u2 n ique sou rce of n itrogen and carbon. Key words: rum en m icroo rgan ism ; m eth ion ine; A ST; AL T; free am ino acid ,,,, 4,,, (J ICST ),, ; 5 300, 2 200, ISSN X, CN gX16, 160 g, : 52-62, : 4721BM, : g :,, : ; ; ; ; ; : ; : 26 : : (029) : (029) E2m ail: bulletin@m s. isw c. ac. cn : h ttp: ggwww. isw c. ac. cn
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