Iimveliso

Imibuzo ibisoloko iphakanyiswa kutshanje: 1. Kutheni uvavanyo lwe-ONPG lungasetyenziselwa ukwahlula ukuvutshelwa kwelactose okulibaziseke?2. Kutheni umgangatho wesizwe uchaza ukuba kuyimfuneko ukusebenzisa i-3% ye-sodium chloride trisaccharide iron (okanye i-trisaccharide iron) yovavanyo lwe-ONPG?3. Kwi-Vibrio parahaemolyticus, xa uqhuba uvavanyo lwe-OPNG, kutheni i-toluene kufuneka yongezwe ngokwe-dropwise ngokomgangatho?Yintoni umsebenzi?

Inkampani yethu iphonononge ulwazi oluninzi, yalushwankathela, kwaye yabelana ngalo nawe ngezantsi:

Umgaqo: Igama lesiTshayina le-ONPG yi-o-nitrobenzene-β-D-galactopyranoside.I-ONPG inokuthi ifakwe i-hydrolyzed ibe yi-galactose kunye ne-yellow o-nitrophenol (ONP) nge-β-galactosidase, ngoko ke umsebenzi we-β-galactosidase unokubonwa ngokutshintshwa kombala wenkcubeko.

ILactose yiswekile ekufuneka ibonwe ziintsholongwane ezininzi.I-metabolism yayo idinga i-enzymes ezimbini, enye i-cell permease, i-lactose ingena kwiiseli phantsi kwesenzo se-permease;enye yi-β-galactosidase, eyenza i-lactose i-hydrolyze ibe yigalactose.ILactose kunye neglucose.I-β-Galactosidase inokuthi isebenze ngokuthe ngqo kwi-ONPG ukuze i-hydrolyze ibe yi-galactose kunye ne-o-nitrophenol emthubi (ONP).Ingenziwa nge-24h, nangona i-lactose ilibazisekile i-fermenters.Ngoko ke, ichaza iziphumo zokujonga inkcubeko yokukha i-1 kwi-agar slant kwaye ifakwe kwisangqa esipheleleyo kwi-36 ° C kwi-1-3h kunye ne-24h.Ukuba i-β-galactosidase iveliswa, iya kuba phuzi kwi-1-3h, ukuba akukho enzyme enjalo, ayiyi kutshintsha umbala kwi-24h.

Ngokwale enzymes zimbini zingasentla, ii-microorganisms zinokwahlulwa ngokwezi ndidi zilandelayo:

I-1 lactose-fermenting (iiyure ze-18-24) ibhaktheriya ene-permease kunye ne-β-galactosidase P + G +;

I-2 i-lactose fermenters ilibazisekile (ithatha ixesha elide kuneeyure ze-24) ayinayo i-permease kodwa ine-galactosidase: P- G+.

I-3 I-non-lactose fermenters ayinayo zombini i-permease kunye ne-galactosidase: P- G-.

Uvavanyo lwe-ONPG lunokusetyenziswa ukwahlula intsholongwane ye-lactose-lag-fermenting (P-G+) kwiibhaktheriya zelactose ezingavundisi (PG-), ezifana nezi:

1 Ukwahlula i-lactose fermenters kade (P- G+) kwi-non-lactose fermenters (P- G-).

(a) I-Citrobacter (+) kunye ne-Salmonella arizonae (+) esuka kwi-Salmonella (-).

(b) Escherichia coli (+) evela kuShigella sonnei (-).

Kutheni le nto uvavanyo lwe-ONPG lwenziwa kusetyenziswa inkcubeko yobusuku kwi-3% ye-sodium chloride ferric trisaccharide (iron trisaccharide)?Inkampani yethu iye yabonisana nolwazi oluninzi, kodwa akukho ngxelo icacileyo.Kuphela kwiwebhusayithi ye-FDA, kubhaliwe ukuba "Iinkcubeko ze-inoculate kufuneka zivavanywe kwi-slants ye-sugar iron agar slants kwaye ifukamele i-18 h kwi-37 ° C (okanye enye iqondo lokushisa elifanelekileyo, ukuba liyafuneka) . Isondlo (okanye enye) i-agar slants equlethe i-1.0 I-% lactose nayo ingasetyenziswa."kuthetha: iibhaktheriya zokuvavanya zifakwe kwi-iron trisaccharide medium kwaye zikhuliswe kwi-37 ° C kwi-18h.I-agar slants (okanye enye) i-medium ene-1% ye-lactose nayo iyamkeleka.Ngoko ke, kucingelwa ukuba i-iron medium ye-trisaccharide iqukethe i-lactose.Emva kokukhula ngobusuku, iibhaktheriya zenze i-β-galactosidase elungileyo esebenzayo.Ukusebenzisa iibhaktheriya ezinjalo, i-ONPG inokubola nge-β-galactosidase ngokukhawuleza.Into yovavanyo ikhawuleza kwaye ibonakaliswe ngcono.Ukongeza, ukongezwa kwe-dropwise ye-toluene kunye nebhafu yamanzi kwi-5 min konke ukwenzela ukukhulula ngokupheleleyo i-β-galactosidase.