ncaa Leto 9 2 1-4 © Sgr 25 Differential patterns of constitutive intracellular laccases of the vegetative phase of Pleurotus species M. Téllez-Télie'?,C, Sinchez', O. Loera? & G, Diaz-Godinee!* "ara of Biotechnology. Rerearch Core or BulogicelScionces, Uni datdna de Tae, Apertads pst 19. CP 9000, Fleas Mesto ‘ttc en Ciencias Bilger, Universit tuna de lca, Mésico Dept, Bienen, Unbersad tama Metropolitan eoptan, Aso DE ‘Air for ecrepondnce (Fes 450268481583. Eoma eect aatsos) ‘ey words eases, myeliom, Pons, yoga Abarat ‘The consiutv intraclass actity often ais of ewer pp termini in ad by ‘amopams ung dilrent strats, Diferene ke the vir ste mere curd between al he ‘uns however ymogrmpateras were ony sil or stra vthin same specie independ a Ofte thee subsate(2dipethoxyphenel, pain or tain) sed Thediewnce rd i he ‘uber and postions ofthe bom inthe pl ug hat ese symograms can be wed to dire ‘pete oct organ. lowodction lke produced by other pes of fing, haces, iets and pln Te fan hse enzymes ma Laces (pptenok: oxygnaoxidorductars; EC aso Be sald in the sap clay vom 111032) “ae” coppercomaining enzymcs that formation of fut bods, cons pipnenaton, ‘atae the oxen of pheobe ssrate by gn degradation, ptogenes and devon. coupling ito the reduction of O; to water Lac: ton of pollutants (Gal wl 2003) Stubs ‘886, manginew-peonidass and liga poroud- op Iss nea have mainly focused on fer Belong to We ensymate comple that extaclolr eduction wing cultures developed ‘la lgnin and are lid as pnoloié- on gu nium (Palos al 2000), ening ter Lares are seed in multiple isoforms, pargeaton and then chracersaton of the Spending mainly on the fungal species and ej (Garden ota. 1995: Marques de Sou {nvronmentl ondons (Giardina ct af 1999). on & Peralta 201). Howene,Icemse activites ‘Thee enayes have been found in whitezot thin meals from diferent Pwo sain ung sch as Plearons pei wich re oen developed om igi has not een ste se fa china on ligninestose.substtes (eg In the work, worm puter of conic ‘maze, barkey or wbeat stan). Plorotur eae: ntrasar sees mee determine by 2mm ‘ais 4 mushroom which 1 thie only to Agar grams for teh Plan sins own oh on om higora (ampigon) and Lente caoies nda agurtnevm. Alera onan (hitko) "in tems of the guantiyprodemd capacity om dileentsbsrates as heed or sound the world (Sinchee 2009). Lactse are ery etc.92 Mate and methods en stains of Pewonus nee: Pores SITS Pos), Pomme 8537 (P03), Poste fa $8052 (Post), Poser 201218 PoP) and ‘Posreans 20121 (Pos) were om the ATCC Eallston, Powe 326 (P28) ma ro the NRRL. pumonar 38 (Pet) and noe paras 7 (Ppt) mee fom the Chine Univer ‘ity of Hong Kong Colleton and 7 eds (PA) nd Fcomiopie (Ps) fom the Universidad da Surde Argentina Colton Iocan sere a tars omits ‘An incaltn was taken from the periphery of Colonies growing on potat\denrve pi" at 38°C for dam, by ing serie cork borer (mm diam) The metal pug. vas plod (aventim facing down) onthe ene a he ele ‘ee mediua. In all experinent, welled ‘sons grown on Pe shes ontining ach ‘carbon source and rnneral sake were ad (Sinetez”& Viniegr-Gonzas 1396) A the "rin ere grown a 25°C, eee for striae of Pipuianares wich were grown at 28°C. The ell exacts were obtained from ayes scape fom the surface of cach colony for en stain Fash mveelum (between 92 and 568 ww of mista) wus grou ng 2 Use Sue pine, whch tld water was oe {ot Tor ea 01mg sample and then centr St 200n>ep for TO min at 2°C, spereatint tre collet und wid Brym aisays Laces activity was asaya ute 26-imeth- fxypienel (DMP) praising ot oliine as ‘nbstes, Tor DMP the aust miu com tained 980 sustate (2 aM DMP in Os Phosphate bur. pH 6.0) and 30 call extract, Which were incated a1'39°C for 1S min. ‘ration of BMDP was flows hy sbsorancs incre 468 nm, For thon ass with pa Sine, the reaction sntre contained 950 substi (10m of panting in Om phos: Dat bute, ft 6) and 8p eal exact, nc fit at 39°C for 1S.mio. Oxidation of "nine wi flowed ya increase in aber ‘ney at 4 pm, Final, when ing oon Subse, he assay. miMtre comaied 950 fubstite (@ mar nthe in 1 a4 acetate bal fer, ft 3.7) ap el extrac, sete at SD%C foe 1S min. Oxisaton of tolidne bas Tetlowed by an icra in absorb "at (20 am. One enzymatic uit (U) of lease ativ= Ay Ws done asthe amount of snyoe which fuse an icra of Tun of abso pe {Bie reaction ature The Ieee soi at exposed U erg dry bom. Pre cosituiveiaraceurlcese activity vas also detected through zymograms sing the Io tehngue SDS-PAGE (Lari 170, ‘The zonaing sel comaiged 100 ¢ acme spd 27g biryani The stacking pt con- tained A gaeramide nd 27g baseyamide {Bich eel etact 30 l aprox) was mito with Sample ble withoot «reducing gent he dle fie bonds Without heating. the apes were ‘placed in gals Chicknes 13 rm) of Mlni-Pro- {ean IT deccophoress sy (Bead) ad then 180-¥ was applied for 1125 h Aor the ‘ecwophoress, gels were mashed wih deonizd Setter on am orbital shaker (2030 rpm) for S's, water mas changed every hour 1 Feene ‘SOS. Fal" the gs oe ncated at oom temparatre in proper sbsteate slurs (DMP, rnin of atbuive, pepe os mentions ‘ove. Laceae ati Bans appeared om the ft by Me exation of the substats ater ppt (2h Rens an dacuion “Table 1 shows he consituveintacellar = ‘ne atvgy of ten sine of Plott on Lédimethowphenal, pansidine ot cxoldine Stine Port and Poe showed the lowest 90 the highest cee activity on all the subsrats, ‘espe. Scher & Vinra-Gonzs 1996) found tht Pot bad about ld ager aa Imclgrowth val om agar and bout double Productivity at pt sale mshroom proton (023 yh and 20 kg fre musiroom per Ae whet saw per day respectively) thn Peerr suo, nr say f pay me Fan mat yen ua ao 3 2a ran tro ‘aso be es rin» sn A #5 Se Du ee ad Wy ANOVA aS Det (G'S ou, Rams put ea SOS pr pea (124 yo and 11 Ag fh mushroom per ‘beat aw er dy. spective. he eset ‘Sic, stmint Post, Pose. apd Pee had Wie lacs acti on DMP and oie and Bid ako abowed peso lw prods pilot production plant (ata ot shown. Thee Tule might ugetthat hos avai showing igh productioy tlt seas alo proud find sted re amounts of acs ito the ‘eam (8 Po. Masur (197 reported ha excel lar asses ar inlet In degradation of Hein ‘or cimination of pina oi hat spe Sig. beste “degrstion Then higher mount of extrac crys in the medi Shane the mobsrateivaion, ering the Taal growth rate and prodeivity. Howes the stuins Po3, Por, P27 and Ppl3t di fo show ay coreaon beeen lca sett nd producti lot poduston plat The Straim mipht be moe sete tothe row onions having more variable pte of Gee Frees | ies ince cess Hom cll oa of teh Phra sais reveled wth) DMP ap so State All sai of Piet showed ln ase bands i the same postion on thee. However, the lataclar asi was Sees (Tab, sagessing tha thse deren ht be senate de fo ili exresion of he oform prods Mansur tat (087) tepid thatthe incl nays ae lavahed in cellu process, morpbogsness and Piment production” Pf and Pharos rata Sunn showed the same ssmogreplc pattern. ic might suggest phylogenete relationship Icon sich strain i fe Eger (1979) ltd Sain of Pero rw Flea, USA. tht was Sale P. “fords and ate cls as Paro fasta “var fons Poe peoced Unigue cease band wth a diferent mala eight when compared to bans fom otber spe See Dpptst and Ppl2? precated. the same Fyingtuphepaten, showing two cese bands Oo lower molest weit than those ofthe re Sf the stain The panindinvendiing and ‘okicineoniding atvites were similar tO ‘howe observed when DMP wu ied at stale (Figue 2) These ress sass dt the sme Irforms ond all the subtits, athoveh et. 2ymagso etyo t Pls ‘tin ag chanel me= with similar enzymati civ. On the other und, 2ymogrm pros fr all substrates so ‘Sst that eae forms were psi Tor each fesse This sei agree with Aran Cees ‘hal (208) who sed mogram patter for “renting pstogenctealy reste specs of Trametes Pec (2000) a0d Moet G99) found that the number and pe othe Indice! lana informs depend on the cule ‘adi ad growth coodtns. I this research, the strains wee grown on nomindctrextie Inedism, whch id vor the change ofthe ses presion. puters Those dis are important to inrtse our owes on te hayes that ae preset hee organs. Sich ight be produced in great amount oF Smnnccelos sutstrats. The difernces b+ ‘ceva in nuber and posion offers in the fl suggests that lacase zmograms mig ‘ty to lees spe of ts ges Ackawutedgement We thane the Mexican Coun of Seience and Technology {CORACYT) Tor sapporing his fear (ope No. 47390. M, Teller ‘ies nupponed by a CONACYT ‘sholaship {irre daring ti work aay Gate § Rec Sra D nt hana naa re et ‘Tmt pen itn es ‘cut Pm Gy Sen A Foti 8, £89 ¥. "Sno the bead of ceophage TH. More 2 se tle Sa €or Ga) ncn a hse he arty ‘Cons Soma 0199) Stub wm cy a peed ‘Un ne gots pr Peer al hin cy Foor WS Spo ome i Boo proce snes af arta cnet th ta Soi Bano maha mee Sw Br Tee ay! esi Row ot Hoe