Mushure memazuva makumi maviri nemashanu ekudzikisa static incubation pa28°C, laccase kubva ku *Pleurotus ostreatus* NRC620 yakaratidza kushanda kwepamusoro-soro mune fungi culture medium. pH yakakwana uye tembiricha yeiyi enzyme yaive 3.0 ne70°C, zvichiteerana. Mushure memaawa maviri ekudzikisa pa40°C ne50°C, kushanda kwe enzyme kwakaramba kuri 68.33% uye 59.61%, zvichiteerana. Mushure memaawa maviri ekudzikisa mu citrate-phosphate buffer (pH 7.0), kushanda kwe enzyme kwakaramba kuri pa100%. Kuwedzerwa kwe10 mM MgSO₄ neCuSO₄ kwakawedzera kushanda kwe enzyme neinenge 21% ne35%, zvichiteerana, nepo NaCl, MnCl₂, KCl, uye CaCl₂ zvakadzivisa kushanda kwe enzyme. Kushandisa ABTS se substrate, kinetic parameters (Km neVmax) ye *Pleurotus ostreatus* NRC 620 laccase yaive 1.99 mM uye 16,217 μmol min−1 L−1, zvichiteerana. Kurapa nema enzymes emuto weapuro kwakaderedza zvakanyanya pH uye viscosity, uye kudzikira uku kwakabatana nekuwedzera kwenguva yekuchengetera. Kurapa nema laccase kwakakonzera kudzikira kudiki kwehuwandu hwe phenolic hwemuto weapuro, asi hapana kudzikira kwekushanda kwe antioxidant kwakaonekwa.
Mumakore achangopfuura, vaongorori vakanyanya kutarisa pakushandiswa kwebiotechnology yakasvibirira muindasitiri yechikafu. Laccase ndeimwe yemaenzyme anobatsira zvikuru muindasitiri yechikafu, ichiwana mashandisirwo munzvimbo dzakadai sekugadzira muto, kubika, kugadzirisa waini, uye kuvandudza hunhu hwe organoleptic hwezvigadzirwa zvechikafu.1Zvirimwa zvakawanda zvepamusoro uye tupukanana tunoburitsa laccase,2uye howa dzakadai se deuteromycetes, ascomycetes, uye basidiomycetes dzinogonawo kugadzira laccase.3Laccase (EC 1.10.3.2) iblue oxidase inoderedza molecular oxygen kuita mvura ichishandisa system ine maatomu matatu akasiyana emhangura, nokudaro ichioxidation yema phenolic compounds akasiyana-siyana uye aromatic amines. Munguva yekugadzira muto wemichero nemiriwo, enzyme nenonenzyme browning inyaya dzakakosha.4Sezvo zvinhu izvi zvichikanganisa ruvara, kunaka, uye kunhuhwirira kwemuto wacho, zvinofanira kubviswa.5
Pamichero yese, maapuro ndiwo anodyiwa zvakanyanya pasi rose uye muEuropean Union. Muna 2019, kugadzirwa kwemaapuro kwaive kwechitatu pasi rose, kuchidarika matani mamiriyoni makumi masere nemanomwe.6Maapuro ane ma phenolic compounds akawanda, anosanganisira flavonoids nema phenolic acids akadai se caffeic acid uye chlorogenic acid.7Nekuti muto weapuro unowanzo nwiwa uri muchimiro chawo chakajeka, inenge 50% kusvika 90% yezvinhu zve phenolic zvinorasika panguva yekusefa.8Nhasi uno, vatengi vanowanzo sarudza zvigadzirwa zvisina kugadzirwa zvakanyanya, zvakaita semuto weapuro une makore akawanda une polyphenol yakawanda. Zvisinei, nekuda kwehuwandu hwawo hwakawanda hwe phenolic, rudzi urwu rwemuto weapuro runonyanya kusviba uye kusviba.9Matekinoroji akasiyana-siyana, anosanganisira nzira dzekurapa kupisa dzakadai se pasteurization pa 60–90°C, anoshandiswa kuderedza kana kudzivirira kusviba kwemuto weapuro.10Zvisinei, maererano nekutsvakurudza kwakaitwa naSauceda-Gálvez11, kugadziriswa kwekupisa kunogona kuparadza makemikari anoshanduka-shanduka uye kukanganisa hunhu hwemuto weapuro. Dzimwe nzira dzekugadzirisa kupisa dzinosanganisira supercritical carbon dioxide, ultraviolet mwaranzi, ultrasound, high hydrostatic pressure, kana high-pressure homogenization.12Kushanda zvakanaka kwetekinoroji idzi uye goho remuto wemichero yakakodzera zvinoenderana nematanho anoshandiswa uye hunhu hwechigadzirwa. Kushandiswa kwazvo kwakapararira kunogumira pamitengo yakakwira, mhedzisiro yakaipa pamhando yezvimwe zvigadzirwa zvekudya, kana kusashanda zvakanaka kwema enzyme.13,14
Laccase inogona kushandiswa kusimbisa uye kujekesa muto wemichero.15Gökmen nevamwe.16Ndinokurudzira kushandisa laccase pakujekesa muto wemichero nekuti inobvisa zvinobudirira phenolic compounds nekushandura kuita mapolymers kana oligomers anobviswa nyore nyore nechero ultrafiltration membrane, zvichiita kuti muto weapuro urambe uine ruvara rwakasimba uye kujeka kwemavhiki matanhatu pa50°C. *Trichoderma* laccase yakacheneswa yakamiswa paalumina beads uye yakashandiswa kubvisa zvine hunhu husina kunaka hunokonzerwa nekusvibiswa kwemuto weapuro nehutachiona.17
Inenge 80-90% yezvinhu zvinonyunguduka zvemuto weapuro zvinoumbwa nemaester nemaaldehydes, izvo zvinopa kunhuhwirira kwakasiyana mumuto wacho.18Laccase kubva ku *Trametes versicolor* yakamiswa kufamba nerutsigiro rwakachipa rwakagadzirwa nefaibha yechisikigo kubva kumagoko ekokonati madiki kuti iwedzere muto weapuro.19Zvidzidzo zvekare zvakaongorora kugadzikana kwemuto weapuro (ruvara uye kusviba) uchishandisa nzira dzisina maenzayimu kana kuti nzira dzekumisa kufamba, kana pamwe chete ne ultrafiltration.5,19Zvisinei, mhedzisiro yemafungus efungus pahunhu hwefizikisi yemuto weapuro panguva yekuchengetwa haisati yanyatsojeka. Nokudaro, chinangwa chechidzidzo ichi chaive chekuongorora nekuyedza shanduko muhunhu hwefizikisi, huwandu hwephenolic compound, uye basa reantioxidant remuto weapuro mushure mekurapwa nemafungus efungus uye kuchengetwa mufiriji kwemavhiki maviri. Mafungus ane kugona kwe oxidize phenolic compounds, izvo zvinoita kuti ave netariro yekushandiswa mumabasa akasiyana-siyana emaindasitiri, kusanganisira kujekesa muto. Chidzidzo ichi chakaongorora mafungus kubva ku *Pleurotus ostreatus* NRC 620, achitarisa mamiriro akakodzera ekushanda kwawo uye kushanda kwawo mukujekesa muto. Kunyange zvazvo tsvakiridzo pamusoro pehowa dze oyster (P. ostreatus NRC 620) ichiri shoma, zvidzidzo zvekare zvakaongorora ma enzymes kubva kunzvimbo dzakasiyana-siyana dzefungus, dzakadai seTrametes versicolor neGanoderma lucidum. Chinangwa chechidzidzo ichi chaive chekuongorora kushandiswa kunogona kuitwa kweiyi enzyme muindasitiri yechikafu uye kuratidza hunhu hwayo hwakasiyana, kunyanya pH yayo yakanaka uye tembiricha.
2,2′-Azooxybis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) yakatengwa kubva kuSigma-Aldrich (Canada). Mamwe ma reagents ese aive emhando yekuongorora.
Nzvimbo yeNational Research Center's Microbial Culture Collection Center yakawana mhando inozivikanwa yehowa re oyster NRC620. Mushure mekurimwa kwembeu, mhando iyi yakachengetwa pamiti ye potato dextrose agar pa 4°C. Nzira yekugadzira inoculum yaive seinotevera: mycelium ine mazuva gumi, yakagadzirwa zvizere, yakaiswa pa potato dextrose agar plates ndokuiswa pa 28°C. Mushure memazuva gumi, mabhuroko matatu e 12-mm-diameter mycelial akabviswa kubva mu agar media achishandisa punch yesimbi isina tsvina ndokuiswa mu 250-mL Erlenmeyer flasks nema cotton plugs ane 50 mL ye sterilized culture medium (pH 5.0, sezvakatsanangurwa naOthman et al.20). Midziyo yacho yakachengetwa pa28°C kwemazuva gumi nemasere. Midziyo yacho yakasefetwa kuburikidza nepepa rekusefa reWhatman No. 1, uye supernatant yakabuda yakashanda senzvimbo inobva enzyme.
Kushanda kweLaccase kwakaongororwa uchishandisa ABTS se substrate. Musanganiswa we reaction (2 mL) waiva ne500 μL ye0.3 mM ABTS (yakanyungudutswa mu 0.1 M sodium citrate buffer, pH 4.5) uye huwandu hunodiwa hwe enzyme sample hwakasanganiswa nemvura yakasanganiswa.21,22Tichifunga kuti laccase inogona kuoxida ABTS patembiricha yemumba (28 °C ± 2), oxidation yeABTS yakayerwa nekuyera kuwedzera kwekunyudzwa kwemvura pa420 nm (ε420= 36,000 cm-1 M -1) uchishandisa Agilent Carry-100 UV spectrophotometer. Chiyero chimwe chete chekushanda kwelaccase chaidiwa kuti chioxidize 1 μmol ABTS paminiti. Kuwanda kwemapuroteni kwakaongororwa nenzira yeBradford uchishandisa bovine serum albumin senzira yekudzora mukati.23,24
Mushure mekuwana enzyme kubva kurudzi rwehowa hweoyster NRC 620, basa rayo rakayerwa panguva dzakasiyana dzekurima kwemazuva makumi maviri nemashanu pasi pemamiriro ekunze asina kusimba pa28 °C.
Kuti tidzidze kuti tembiricha inokonzereswa sei pakushanda kwe laccase, kuyedza kwakaitwa muchikamu chetembiricha kubva pa20 kusvika 90 °C. Vasati vawedzera enzyme uye vatanga kuita reaction, buffer (0.1 M sodium citrate, pH 4.5) uye substrate (ABTS) zvakasanganiswa uye zvakaiswa mu incubation kwemaminitsi mashanu patembiricha dzakasiyana. Kugadzikana kwe enzyme kwakaongororwa ne incubation mu 0.05 M sodium phosphate buffer (pH 7.0) pa 40, 50, 60, uye 70 °C kwemaawa maviri, zvichiteerana. Basa rekusara rakazoongororwa uchishandisa substrate yeABTS.
Mhedzisiro yepH pakushanda kwe laccase yakaongororwa uchishandisa ABTS se substrate mu 0.1 M citrate-phosphate buffers ine pH range ye2.5 kusvika 7.0. Mhinduro ye enzyme yakaiswa pa40°C kwemaawa maviri mu 0.1 M citrate uye Tris buffers (pH 3, 4, 6, uye 7) kuti iongorore kugadzikana kwepH. Basa rekusara neABTS se substrate rakaverengerwa mushure mekuiswa.
Laccase yakachengetwa kwemaminitsi gumi mu sodium phosphate buffer (0.05 M, pH 7.0) ine ma ions akasiyana-siyana esimbi (Mg2+, Cu2+, Co2+, Ca2+, Zn2+, K+, Na+, uye Mn2+) pahuwandu hwe2.5 mM uye 10 mM, zvichiteerana. Substrate (ABTS) yakawedzerwa kuti itange reaction, uye relative activity yakaongororwa.
Kuoxidation kweABTS ne laccase pahuwandu hwakasiyana (0.025–3 mM) kwakayerwa pa pH 4.5 kuti kuonekwe ma parameter e kinetic (Vmax ne Km).zvisingaperiyeMichaelis-Menten equation yakaverengerwa uchishandisa Lineweaver-Burk plot, iyo inonyora reciprocal ye reaction rate sebasa re substrate concentration. Kinetic constants yakaverengerwa kubva kuLineweaver-Burk plot uchishandisa GraphPad Prism version 6.01 software.
Mushure mekusuka maapuro nemvura yepombi zvakanaka, akachekwa nepakati ndokuiswa juicer uchishandisa muchina weBraun MP80 apple juicer (wakagadzirwa muGermany). Muto wacho wakaseferwa kuburikidza nejira rechiziro mana. Hapana ma enzyme akawedzerwa kuboka rekutonga, nepo 2.0% laccase (huwandu hwakanyanya kuyedzwa) yakawedzerwa kumuto weapuro wakagadzirwa, uyo wakazochengetwa pa4°C kwemavhiki maviri.
Titratable acidity (TA) uye pH zvakaongororwa zvichienderana nenzira yaBoulton et.al.27. PH yemuenzaniso wega wega yakayerwa uchishandisa digital pH meter (JENWAY 3510 pH meter). Titratable acidity (TA) yakaverengerwa zvichibva pa malic acid uchishandisa fomura inotevera.
Apo V naC zviri vhoriyamu (mL) uye huwandu (0.1 mol/L) ye sodium hydroxide solution inoshandiswa mu titration, zvichiteerana. K ndiyo malic acid conversion coefficient, yakaenzana ne 0.067, uye W ndiyo huremu (g) hwemuto weapuro.
Zvinhu zvose zvinonyunguduka (TDS) huwandu hwemasampuli ese emuto hwakaongororwa uchishandisa PAL-1 pocket refractometer (ATAGO, Tokyo, Japan). Mushure mekuyerwa kwega kwega, lenzi yemaziso yakasukurudzwa nemvura yakabviswa maion, uye sampuro yega yega yemuto weapuro yakaedzwa katatu. Kukosha kwesampuro yega yega kwakaverengerwa neavhareji yezviyero zvitatu. Avhareji ± standard deviation yesampuro yega yega yemuto weapuro yakaverengerwawo neavhareji yemhedzisiro iyi.
Kupenya kwemvura yemuto weapuro kwakaongororwa uchishandisa rotational viscometer (RV, Rheotest 2, Germany). Muenzaniso wakaiswa mukati me "S2" cylinder yeviscometer. Kupenya kwakajeka kwaimirirwa nekutsveyama kwe shear stress vs shear rate curve, iyo yakaverengerwa kubva pa shear stress uye ma curves anoenderana pa shear rates dzakasiyana (kubva pa 1.00 kusvika 437.4 s⁻¹). Fomura yekuverenga viscosity inooneka ndeiyi inotevera:
Apo η iri pachena viscosity (cP), τ iri shear stress (dyn/cm²), γ iri shear rate (sec⁻¹), uye (τ) inoverengwa uchishandisa torque (α) uye cylinder (Z) values uchishandisa fomura inotevera: τ = Z . α.
Nhamba yebrowning yakatarwa zvichienderana nenzira yaMeidav etal.29. Sampuro yemuto we10-ml yakatenderedzwa ne centrifuge pa 2750 xg kwemaminitsi gumi. 5 ml yemuto we supernatant yakasanganiswa ne 5 ml ye 95% ethanol. Kunyudzwa kwemusanganiswa kwakayerwa pa 420 nm uchishandisa Shimadzu UV spectrophotometer (UV-1601 PC).
Huwandu hwe phenolic content (TPC) hwakaongororwa neruvara rwemavara uchishandisa Folin-Ciocalteu reagent sezvakatsanangurwa naBoulton et al.[27]]. Kavha yakajairika yegallic acid yakagadzirwa kuti ive nehuwandu hunobva pa0 kusvika 500 mg/L (r²= 0.997). Mhedzisiro yacho inoratidzwa se gallic acid equivalents (mg GAE/mL).
Wedzera 125 μL yemvura yakasanganiswa uye 2850 μL yeFRAP solution ku25 μL yemuto weapuro wosiya musanganiswa wacho murima kwenguva yakareba.30min. Wobva wayera absorbance pa593 nm uchishandisa Shimadzu UV spectrophotometer (UV-1601 PC). FRAP reagent yakagadzirwa nekusanganisa 300 mM acetate buffer (pH 3.6), 20 mM iron(III) chloride, uye 10 mM 2,4,6-tris(2-pyridyl)triazine (TPTZ) (yakanyungudutswa mu40 mM HCl) muchiyero che10:1:1. Mutsetse wakajairwa wakagadzirwa uchishandisa Trolox semupimo (R²= 0.999), uye mhedzisiro yacho inoratidzwa se μM Trolox/mL.
Kushanda kweantioxidant kwemajusi akarapwa neasina kurapwa kwakaongororwa uchishandisa nzira yeDPPH kuti vaongorore kugona kwavo kubvisa mafree radicals eDPPH.31Ma microliters gumi emuto akasanganiswa ne 1 ml ye DPPH solution (100 μM) mu methanol. Mushure mekuita kwekuita murima kwemaminitsi makumi matatu, kunyudzwa kwemusanganiswa kwakayerwa pa 517 nm uchishandisa Shimadzu UV spectrophotometer (UV-1601 PC). Mhedzisiro yacho yakatsanangurwa se trolox equivalents (μM trolox/ml) zvichibva pa calibration curve (R2= 0.990).
Data rakawanikwa rakaratidza kuti kugadzirwa kwakanyanya kwehowa dzeoyster muNRC 620 kwakaonekwa muhowa weoyster mushroom weNRC 620 pakupera kwezuva rechi18 rekuvirisa, kusvika pakushanda kwe1302 U/L. Izvi zvakashanda sehwaro hwekuona nguva yakakodzera yekurima howa dzehowa dzehowa (Mufananidzo 1). Kunyangwe kugadzirwa kwema enzyme kwakawedzera nekuwedzera kwenguva yekurima, mwero wekuwedzera wakanga usina kuenderana zvakananga nenguva yekurima; mushure memazuva makumi maviri nerimwe, kushanda kwema enzyme kwakawedzera ne90 U/L chete (kusvika 1390 U/L). Saka, mazuva gumi nemasere akasarudzwa senguva yakakodzera yekurima kuti pave nekuenzanisa goho rezvigadzirwa nehupfumi hwenguva yekurima yakawedzerwa.
Mhedzisiro yenguva yekurima pagoho re laccase muPleurotus ostreatus NRC 620. Mabhuroko matatu (12 mm) efungus mycelial akabayiwa mu 50 ml ye sterile medium ndokuzorimwa pa 28 °C kwenguva dzakasiyana.
Zvichienderana nedzimwe ongororo, zvatakawana zvinoratidza kuti nguva yakakodzera yekurima kuti fungus ibudise laccase yakawanda inogona kunge iri pakati pemazuva manomwe kusvika makumi matatu nematanhatu.32Sekureva kwaEzike nevamwe.33, *Trametes polyzona* WRF03 yakagadzira huwandu hwakawanda hwe laccase pakupera kwezuva rechipfumbamwe rekuvirisa, iine basa rakati re 1637 U/mg protein. Uyezve, Othman et al.34vakawana kuti *Trichoderma harzianum* S7113 yaiburitsa laccase yakawanda pazuva rechishanu rekurimwa. Kukura kwelaccase kwakasvika padanho repamusoro pazuva rechigumi nemana uye kwakazodzikira zvishoma nezvishoma.34Kunyangwe kuburitswa kwema enzymes kuchigonawo kuitika panguva yekukura kukuru, kunowanzo svika padanho repakati uye kunokonzerwa nekushandiswa kwekabhoni kana nitrogen.34,35
Kunyangwe laccase kubva kuPleurotus ostreatus NRC 620 yakaratidza kushanda kwakanyanya pamusoro pekupisa kwakakura kubva pa50°C kusvika 80°C, ichisvika pakuita kwakanyanya (69–98%), kushanda kwayo kwakanyanya kwakaonekwa pa70°C (Mufananidzo 2a). Kunze kwekupisa uku, kushanda kwema enzyme kwakaderera pa70°C. Mhedzisiro iyi inoratidza kuti enzyme inoshanda pakupisa kwakanyanya, zvichida nekuti kupisa kwakanyanya kunowedzera simba rekuita.
Mhedzisiro yekupisa kwe reaction (a) uye pH (b) pa laccase activity mu *Pleurotus ostreatus* NRC 620. Kupisa kuri pakati pe20 kusvika 90 °C kwakawanikwa nekuisa musanganiswa patembiricha dzakasiyana kwemaminitsi mashanu usati waisa enzyme uye kutanga reaction. Mhedzisiro ye pH pa laccase activity yakaongororwa uchishandisa ABTS se substrate mune mhinduro dzine 0.1 M citrate-phosphate buffer pamusoro pe pH range ye2.5 kusvika 7.0.
Sekureva kwaEzike etal.33, tembiricha yakanakisisa ye *Trametes polyzona* WRF03 laccase i55 °C, inova yakafanana neye *Ganoderma lucidum*laccase36uye yakafanana netembiricha yepamusoro (50 °C) ye *Trametes polyzona* KU-RNW02737laccase . Baldrian38anoti, semamwe masisitimu eenzyme anoparadza lignin, tembiricha yakakodzera yelaccase iri pakati pe50 ne70 °C.
Zvakabuda zvakaratidza kuti enzyme iyi yakaratidza kushanda kwepamusoro pa pH 3.0, ichisvika 94% pa pH 3.5. Zvisinei, yakaramba ichishanda pane pH yakakura kubva pa 2.5 kusvika 7.0 (Mufananidzo 2b). Uyezve, yakaratidza kushanda kwakanyanya mumamiriro eacidic zvichienzaniswa nemamiriro ekunze asina kusimba kana alkaline. Kushanda kwayo kwakaramba kuri 77% kupfuura pH kubva pa 2.5 kusvika 4.5, asi kwakasvika pa 38% chete pa pH 7.0. pH yakanakisisa ye laccase kubva ku *Trametes polyzona* WRF03 yaive 4.533, iyo yakafanana ne pH ye laccases kubva ku *Trametes polyzona* KU-RNW02737, *Trichoderma harzanium* 39, *Pleurotus* sp. 40, uye *Trametes hirsuta* 41. Zvisinei, zvichienderana nekudzidza kwakaitwa naChairin et al.42, pH yakanakisisa ye laccase kubva ku *Polymorpha f. sp.* WR710-1 iri 2.2, nepo pH yakanakisisa ye laccase kubva ku *Polymorpha f. sp.* IBL-04 iri 5.043. Kusungana kwe hydroxide anions (laccase inhibitor) kumaatomu emhangura e T2/T3 laccase kungave kuri chikonzero chekuderera kwekushanda kwe laccase pasi pemamiriro e pH asina kusimba kana alkaline. Izvi zvinogona kukanganisa kutamiswa kwema electron emukati kubva pakati pe T1 kuenda pakati pe T2/T3, nokudarokudzikamisabasa re enzyme23,44
Nekuisa enzyme iyi patembiricha dzakasiyana, zvakaonekwa kuti nguva yekuiswa kwemasero pamwe netembiricha zvakakanganisa kugadzikana kweenzyme. Zvinonyanya kukosha, laccase kubva ku *Trametes polyzona* NRC 620 yakaratidza kugadzikana kwakanyanya pa40℃ ne50℃, ichichengetedza 68.33% ne59.61% yekushanda kwayo kwekutanga, zvichiteerana, mushure memaminitsi 120 (Mufananidzo 3a). Kusiyana neizvi, pasi pemamiriro akafanana (40℃ ne50℃, maminetsi 120), laccase kubva ku *Trametes polyzona* WRF03 yakachengetedza 64.38% ne42.92% yekushanda kwayo, zvichiteerana.33Kusiyana neizvi, kuwedzera kwenguva yekubatidza nekupisa kwakaderedza kugadzikana kwe *Trametes polyzona* NRC 620 laccase; Mushure mekubatidza pa60℃ ne70℃ kwemaminitsi makumi matanhatu, kushanda kwayo kwakaderera kusvika pa39.24% ne1.72%, zvichiteerana (Mufananidzo 3a). Zvichienderana nemigumisiro yekuyedza, laccase kubva ku *Trametes polyzona* WRF03 yakaratidza kugadzikana kwakanyanya pa40 ne50℃ panguva yese yekurapa kwekupisa.33Saizvozvowo, Lueangjaroenkit etal.37uye Chairin etal.42yakashuma kugadzikana kwelaccases kubva kuTrametes polyzona KURNW027 neTrametes polyzona WR710-1 pa50 °C kweawa imwe chete, zvichiteerana. Semushonga unobatsira webiocatalyst unoshanda muminda yakasiyana-siyana yebiotechnology, laccase inofanira kunge yakagadzikana uye ichishanda zvakanaka pakupisa kwakakura.
Kugadzikana kweThermostatic (a) uye kugadzikana kwepH (b) kwelaccase kubva ku *Pleurotus ostreatus* NRC 620. Kugadzikana kweThermostatic kwakaongororwa nekuisa enzyme solution mu 0.05 M sodium phosphate buffer (pH 7.0) pa 40, 50, 60, uye 70 °C kwemaawa maviri, zvichiteerana. Kugadzikana kwepH kwakaongororwa nekuisa enzyme solution mu 0.1 M citrate buffer uye Tris buffer (pH 3, 4, 6, uye 7) pa 40 °C kwemaawa maviri. Basa rekusara rakaverengerwa uchishandisa ABTS se substrate mushure mekuiswa mukati memvura.
Kuti tione mamiriro akakodzera ekushandiswa nekuchengetwa kwema enzyme, takaongorora mhedzisiro ye pH pakugadzikana kwe laccase. Kusangana ne pH dzakasiyana kwakakanganisa zvakanyanya kugadzikana kwechimiro cheprotein, nokudaro zvichikanganisa kugadzikana uye kushanda kwe enzyme molecule. Mhedzisiro yakaratidza kuti enzyme yakanga isina kugadzikana zvakanyanya mumamiriro ezvinhu ane acidic, nepo yakaratidza kugadzikana kuri nani pane pH yakakwira (nzvimbo dzisina kusimba uye dzine alkaline). Pa pH ya7.0, 6.0, 4.0, uye 3.0, mwero wekuchengetedza ma enzyme mushure memaminitsi 120 waive anenge 100%, 62.54%, 52.39%, uye 11.14%, zvichiteerana (Mufananidzo 3b). *Strombus multisus* WRF03 laccase yakaratidza kugadzikana kwakanyanya pane neutral pH values (5.5–6.5) uye kugadzikana kwakaderera pane acidic pH values (pasi pe4.0). Mushure memaminitsi zana nemakumi maviri pa pH ye5.5, 6.0, uye 6.5, mwero wekuchengetedza ma enzyme waive anenge 82%, 100%, uye 93%, zvichiteerana.33Khairin nevamwe.42vakaona kuti laccase kubva kuTrametes polyzona WR710-1 yakanga yakagadzikana mu pH range ye6.0 kusvika 7.0, nepo Sayed nevamwe.45yakaratidza kuti laccase yakanga yakagadzikana zvakanyanya pasi pemamiriro e pH asina kusimba. Zvisinei, laccase kubva kuCerrena unicolor yakaratidzawo kugadzikana pasi pemamiriro e alkaline (pH 9.0)46. Madhaka akaongororwa akaratidza kugadzikana kwakanyanya pamusoro pehuwandu hwakakura hwepH. Izvi zvinogona kuva hunhu hwakakosha pakushandiswa kwemaindasitiri.
Sezvo mamwe maion esimbi aine mhedzisiro yekukurudzira uye yekudzivirira pabasa remaenzyme, mhedzisiro yawo pabasa remaenzyme inofanira kufungwa nezvayo mumaindasitiri. Izvi zvakakosha nekuti maion esimbi ndiwo anowanzo svibisa nharaunda anogona kukanganisa kugadzikana uye kugadzirwa kwemaenzymes ari kunze kwemuviri.47Kuti tiongorore mhedzisiro yemaion akawanda esimbi pa laccase kubva ku *Pleurotus ostreatus* NRC 620, takaita bvunzo dzinoenderana. Sezvakaratidzwa muMufananidzo 4, zvichienderana nerudzi rwesimbi inoshandiswa, kuwedzera huwandu hweion yesimbi kubva pa2.5 mM kusvika pa10 mM kwakakanganisa mashandiro e enzyme. Semuenzaniso,Mg²⁺ , Co²⁺ , Zn²⁺uyeCu²⁺inogona kukurudzira uye kumutsa basa re enzyme, ukuwoNa⁺ , Mn²⁺ , Ca²⁺uyeK⁺yaigona kumisa mashandiro eenzyme. Pahuwandu hwe10 mM, maion eCu²⁺ neMg²⁺ ndiwo ainyanya kumutsa mashandiro elaccase kubva ku *Pleurotus ostreatus* NRC 620, zvichipa mwero wekushanda weinenge 34% ne20%, zvichiteerana. Zvisinei, pahuwandu hwe10 mM, maion eCa²⁺ ndiwo ainyanya kudzivirira laccase, zvichideredza mashandiro eenzyme neinenge 60%.
Mhedzisiro yemaion esimbi pabasa rePleurotus ostreatus NRC 620 laccase. Laccase yakachengetwa kwemaminitsi gumi mu sodium phosphate buffer (0.05 M, pH 7.0) ine maion akasiyana-siyana esimbi pahuwandu hwe2.5 mM uye 10 mM. Mhinduro yacho yakazotangwa nekuwedzera substrate (ABTS), mushure mezvo chiitiko chehukama chakayerwa.
Zvatakawana zvinoenderana nezvakawanikwa nevamwe vanyori vakaona kuti Mg²⁺ naCu²⁺ zvinowedzera kushanda kwe *Trametes polyzona* WRF03³. Castaño nevamwe vakaona kuti laccase kubva ku *Xylaria* sp. inosimudzirwa kusvika pamwero wakati nemaion emhangura (Cu²⁺). Uyezve, Foroutanfar nevamwe.⁴⁹ naSi nevamwe.⁵⁰ vakaita zvidzidzo zvakafanana pama laccases kubva ku *Paraconiothyrium variabile* uye *Trametes pubescens*, zvichiteerana. Nzvimbo yekubatanidza mhangura yerudzi rwechipiri (T2) yeiyi enzyme inogona kuzadzwa neCu²⁺ pahuwandu hwakatarwa, izvo zvinogona kutsanangura kukurudzira kwekushanda kwe laccase pahuwandu hwakakwira hweCu²⁺³⁹. Sezvo fungi chena dzakaora, laccases dziri maoxidases ane maatomu akawanda emhangura, mhedzisiro yemaion emhangura pakushanda kwelaccase yakasiyana uye inotangira pakukurudzira uye kudzivirira kusvika pakusapindirana.⁵¹ Kusiyana neizvi, Zhou nevamwe vake. [52]yakashuma kutiCu²⁺yakadzivisa mashandiro elaccase eTaiwan subterranean termite (Odontotermes formosanus). Zvisinei, laccases dzeCerena sp. HYB07[53]uye Clitocybe maxima[54]hazvina kukanganiswa nemaion emhangura.
Kunyatsoenderana kwesubstrate kwaimirirwa nemakinetic parameters ayo (Km neVmax); kana kubatana kwesubstrate neenzyme kwakasimba, Km value yakaderera uye substrate specificity yakakwira.3,21,55Ma parameter e kinetic (Km neVmax) e laccase kubva ku *Pleurotus ostreatus* NRC 620 akawanikwa achishandisa GraphPad Prism 6.0 software nekunyora Lineweaver-Burk plot (Mufananidzo 5). Pakushandisa ABTS se substrate, mhedzisiro yacho yaive 1.99 mM uye 16217 μmol.min⁻¹ L⁻¹,Elsayed nevamwe.21yakashuma kuti maKm values eABTS oxidation aive 0.1 mM uye 0.064 mM, zvichiteerana, zvichiratidza hukama hwakakura hweLac A neLac B isoenzymes yeABTS. Uyezve, maVmax values aive 0.182 μmol.min⁻¹uye 0.603 μmolmin⁻¹, zvichiteerana. Kukosha kweKm kwakawanikwa kwaive kwakaderera pane kweTrametes polyzona WRF03 (8.66 mM); uyezve, kukosha kwavo kweVmax (1429 mmol min⁻¹) kwaivewopasipakushandisa ABTS se substrate.33 Saizvozvowo, maKm values eLentinus squarrosulus MR13 neTrametes sp. AH28-2 laccase concentrations aive 0.0714 mM uye 0.025 mM, zvichiteerana, uye maVmax values aive 0.0091 mM min−1 uye 0.67 mM min−1 mg−1 (zvichienderana neABTS), zvichiteerana.56,57
Mhedzisiro yehuwandu hweABTS pabasa re laccase kubva ku *Pleurotus ostreatus* NRC 620 yakaongororwa, uye gwaro re Lineweaver-Burk re reciprocal re initial reaction velocity vs ABTS concentration rakarongwa. Maitiro e oxidation eABTS ane huwandu hwakasiyana (0.025–3.0 mM) ye laccase akayerwa pa pH 4.5 kuti aone kinetic parameters (Vmax ne Km). Michaelis-Menten kinetic constants akaverengwa achishandisa Lineweaver-Burk plot ye reciprocal ye reaction velocity vs substrate concentration. Ma kinetic constants akaverengwa kubva pa Lineweaver-Burk plot uchishandisa GraphPad Prism 6.01 software.
Maenzymes echinyakare anotsanangura, akadai sepectinases, anobvisa pectic substances, achideredza viscosity uye turbidity. Anoputsa zvinobudirira mapolysaccharides emukati uye anowanzo shandiswa pamwe chete nemamwe maenzymes, akadai secellulases nehemicellulases, kuti awedzere goho uye kujeka. Zvisinei, mapectinase haanyatsonanga ma phenolic compounds, ayo ari iwo anonyanya kukonzera turbidity uye oxidative browning, kunyanya mumajusi akadai seapuro nemuto wemazambiringa.58Kusiyana neizvi, ma laccase anokonzera oxidation yema phenolic compounds, achiaita polymer kuita mamorekuru makuru, asinganyungudike anogona kubviswa ne sedimentation kana filtration. Iyi nzira haingogadzirisi kujeka chete asi inowedzerawo hupenyu hwesherufu yemuto nekuderedza mukana we oxidative browning inokonzerwa nema phenolic compounds. Uyezve, maitiro ekujekesa akavakirwa pa laccase anogona kuitwa mumamiriro ekugadzirisa akapfava (pH 3.5–5.5, tembiricha 25–40 °C), zvichiita kuti akwanise kushandiswa nemajusi asina kusimba pasina kukanganisa hutano hwavo kana organoleptic.59Zvidzidzo zvakaratidza kuti kurapwa kwepectinase kunogona kujekesa muto mumaawa 1-2, nepo kurapwa kwelaccase kuchiwanzoda nguva yakareba yekuita (maawa 3-6) kuderedza zvachose ma phenolic compounds. Zvisinei, maitiro aya anogona kugadziriswa nekumisa kufamba kwe enzyme kana nekubatanidza laccase nenzira dzekuchenesa michina.60Muchidzidzo ichi, kuongorora enzymes yechinhu chisina kucheneswa kwakaratidza mabasa makuru e laccase ne α-amylase, nepo mabasa e pectinase ne xylanase aive akaderera zvakanyanya, uye mashandiro e cellulase haana kuonekwa. Nokudaro, kudzikira kwe turbidity uye phenolic content kwainyanya kukonzerwa nekushanda kwe laccase, nepo kuchinja kwe viscosity kunogona kunge kuri nekuda kwekushanda kwe amylase.
Tafura 1 inoratidza zviyero zvefizikisi zvemuto weapuro wakasvinwa uye sampuli dzakagadziriswa laccase. Zvakabuda zvakaratidza kuti goho remuto weapuro wakasvinwa (71.59%) raive rakaderera pane remuto wakasvinwa laccase (87.34%). Izvi zvinoenderana nezvakawanikwa naPilnik naOrange.61, avo vakaratidza kuti kushandiswa kwema enzymes mukugadzira michero kunogona kuwedzera goho remuto, kunatsiridza kusefa, uye kuwana muto wemhando yepamusoro, wakajeka wehuwandu. Kuwedzera kwegoho remuto kunonyanya kukonzerwa nekuwedzera kwehuwandu hweshuga inonyungudika mumuto. Munguva ye enzyme hydrolysis yemichero, mesoglea ne pectin mumadziro esero emuto zvinoparadzwa uye zvinoshandurwa kuita zvinhu zvinonyungudika senge shuga dzisina neutral uye acids.62.pH yemuto weapuro wakagadziriswa ne enzyme yaive yakaderera zvakanyanya pane yeboka rekutonga (P < 0.05), uye pH yemapoka ese ari maviri yakawedzera zvakanyanya panguva yekuchengetwa (Tafura 1). Mhedzisiro iyi inoenderana neyaMark et al.63, avo vakataura kuti pH yemuto wemichero yecashew yakaderera mushure mekuchengetwa mushure mekupiswa. Kuora kwepectin uye kuumbwa kwegalacturonic acid mushure mekuchengetwa kwema enzyme zvinogona kukonzera kuwedzera kwepH panguva yekuchengetwa. pH yemasampuli akarapwa nema enzyme yakaramba iri pakati pe4.05 ne4.31 panguva yekuchengetwa, nepo pH yemuto weapuro usina kurapwa yaiva pakati pe4.12 ne4.33.
Huwandu hweacidity (TA) hwesamples dzisina kurapwa nedzakagadziriswa laccase hwakaratidza kudzikira kwemaitiro nekuwedzera kwenguva yekuchengetera (Tafura 1). Kudzikira kweacidity kwakakonzerwa nekushandurwa kwema organic acids kuita makabhohaidhiretsi kana enzyme reactions, pamwe ne oxidation panguva yekuchengetwa kwejusi.64Huwandu hweacidity yemuto weapuro wakagadziriswa uye sampuli dzakarapwa nema enzyme hwakanga hwakaderera pane hwemamwe majusi (muto westrawberry 0.9%, muto weplum 2.2%, muto wekumquat 1.0%, muto weapricot 2.4%, muto weorenji 0.8%), asi hwakafanana nehwemamwe majusi (semuenzaniso, muto wepeya 0.3%).62Kusiyana uku mumuto weapuro usina kurapwa kunogona kukonzerwa nezvinhu zvakasiyana-siyana zvakaita semamiriro ekukura, majini, nguva yekukura, uye nzira dzekugadzirisa.65Kuderera kweacidity yese yekudzora uye laccase-treated apple juice zvinoenderana nemigumisiro yakapihwa naSingh et al.66maererano nekuderera kweacidity yese yemuto weapuro weJin Nuo mushure memazuva makumi manomwe nemana ekuchengetwa. Kune rumwe rutivi, Oshmiansky naWojdylo67Havana kuwana shanduko huru muacidity yemuto weapuro pavaidzidza nezvemhedzisiro yenzira dzechinyakare dzekujekesa.
Mhedzisiro yakaratidzwa muTafura 1 inoratidza kuti huwandu hwese hwezvinhu zvinonyungudika (TSS) hwemuto weapuro wakarapwa nelaccase hwaive hwakakwira kupfuura hwemuenzaniso usina kurapwa. Mhedzisiro iyi inoenderana nezvidzidzo zvakaburitswa.. 68Uyezve, Tafura 1 inoratidza kuti kukosha kweTSS kweboka remuto weapuro waive 9.58 panguva yekutanga uye kwakasvika 11.05 pakupera kwenguva yekuchengetera. Kukosha uku kwakaderera pane kukosha kweTSS kwemuto weapuro mutsva kwakataurwa naHamid et al.. 69(11.2 na11.80, zvichiteerana). Kukosha kweTSS kwemasampuli emuto weapuro wakagadziriswa laccase kwakawedzera zvakanyanya, kutanga kubva pa11.23 kusvika pa12.93 mushure memavhiki maviri ekuchengetwa pa4°C (Tafura 1). Kuwedzera kwakafanana kweTSS panguva yekuchengetwa kwakaonekwawo mumichero yecitrus, maremoni, uye maranjisi anotapira. Kuwedzera kwezvimedu zvinonyungudika zvachose (TSS) panguva yekuchengetwa kunogona kunge kuri nekuda kwehydrolysis yepolysaccharides (starch) kuita monosaccharides (shuga), kuwedzera kwehuwandu nekuda kwekushaya mvura mumuviri, uye kuora kwepectin mumuto kuita soluble solids. Kuwedzera kwezvimedu zvinonyungudika zvachose (TSS) kungangove nekuda kwekuwedzera kweshuga dzinonyungudika, izvo zvinogona kuumbwa nekushandurwa kwepectin kana cellulose kuita soluble sugars nepectin kana cellulase, zvichiteerana, kana nehydrolysis yestarch kuita shuga, sezvakashumwa naHamed et al.69.Mhedzisiro ye laccase pahunhu hwemuto weapuro inogona kuonekwa nemaziso, sezvo muto weapuro wakagadziriswa laccase uchiratidza kuyerera kuri nani uye kusimba kwakaderera pane muto usina kurapwa. Kucherechedzwa uku kwakanyorwa muTafura 1; Kusimba kwemuenzaniso wakagadziriswa enzyme kwaive 1.87 cP, nepo kusimba kwemuenzaniso wekudzora kwaive 2.95 cP. Kudzikira kukuru kwe viscosity uku kungangodaro kuri nekuda kwekukwanisa kwakakwirira kwekubata mvura kwezvinhu zvakaita se pectin uye kuumbwa kwechimiro che network chakabatana.
Muchidzidzo ichi, mhedzisiro yelaccase pabrowning index (BI) yemuto weapuro yakaongororwa nekuyera kunyudzwa kwemuto pa420 nm uchishandisa spectrophotometer. Mhedzisiro yacho inoratidzwa muTafura 1. Munguva yekuchengetwa, BI yemasampuli emuto weapuro mumapoka ese ari maviri akarapwa neasina kurapwa yakaratidza kuwedzera zvishoma nezvishoma. BI inoratidza mwero webrowning uye inogona kushanda sechinhu chakakoshachiratidzo chemaitiro e enzyme neasiri enzymatic browning reactions. Kunyudzwa kwacho kwakawedzera zvakanyanya panguva yekuchengetwa (P < 0.05). Pakupera kwekuchengetwa,A420Kukosha kwemasampuli emuto weapuro mumapoka ekutonga neakarapwa nema enzyme kwakawedzera neinenge 217% uye 121%, zvichiteerana (Tafura 1). Zvakabuda zvinoratidza kuti kurapwa kwema enzyme kunogona kuderedza mwero webrowning neinenge 56%. Zvakabuda kuna Bezerra et al.[19]] zvinoenderana nezvatakawana; Vakashandisa laccase-glutaraldehyde-coconut fiber kuti vagadzire muto weapuro, vachideredza ruvara rwawo rwepakutanga ne61%.
Kunyangwe ma polyphenols ari mumichero ane mhedzisiro yakanaka yekudya uye yekurapa pamuviri wemunhu, anogonawo kuita nemapuroteni, zvichikonzera kusviba kwemvura, kudonha kwemvura, kana kusviba, nokudaro zvichichinja kunaka nekunhuhwirira kwechigadzirwa uye kuderedza nguva yachinosara.71Chinangwa chechidzidzo ichi chaive chekuderedza zvakachengeteka huwandu hwe phenolic compound mumuto weapuro uchishandisa laccase kubva kuPleurotus ostreatus NRC 620. Mhedzisiro yakapihwa muTafura 1 inoratidza kuti huwandu hwe phenolic compound mumuto weapuro wakagadziriswa laccase hwakaderedzwa zvakanyanya usati wachengetwa pa4 °C. Uyezve, huwandu hwe phenolic compound hwakadererawo panguva yekuchengetwa mumienzaniso yese yakadzidzwa (Tafura 1). Tsvagiridzo yakaitwa naSandri et al.72yakaratidza kuti muto weapuro wakagadziriswa nema enzyme unogona kuchengetedza basa rawo rekuchengetedza ma antioxidants uye huwandu hwe phenolic compound. Zvisinei, mhedzisiro yechidzidzo chakaitwa naLettera et al.73zvinoratidza kuti kurapwa kwemuto weorenji nefungus laccase kunogona kuderedza huwandu hwe phenolic compounds mairi kusvika ku45%.
Zvinoratidzwa kuti ma phenolic compounds ane zvinhu zvakaita sekubvisa ma free radicals, kuderedza nekudzima okisijeni mumasero, kutamisa maatomu ehydrogen, uye kupa maerekitironi kuma free radicals, zvichiita kuti ave ma antioxidants ane simba.74Saka, muchidzidzo ichi, nzira dzeDPPH neFRAP dzakashandiswa kuongorora mhedzisiro yelaccase pakushanda kweantioxidant kwemuto weapuro wakachengetwa mufiriji kwemazuva gumi nemana (Tafura 2). Nzira dzese dziri mbiri dzakaratidza kuwedzera kwekushanda kweantioxidant panguva yekuchengetwa, izvo zvinogona kunge zvichikonzerwa nekuwedzera kwemakemikari ephenolic akasununguka kana kuumbwa kwezvigadzirwa zveMaillard reaction (MRPs), nezvigadzirwa zveMaillard reaction zvingangove zviri izvo zvinokonzera kuwedzera kwekushanda kweantioxidant.75Maitiro asina enzyme browning reactions (kusanganisira ascorbic acid degradation, Maillard reactions, uye acid-catalyzed degradation of sugar) anogadzira pigments dzebrown (melanoidins). Zvigadzirwa zvepakati peascorbic acid degradation uye zvigadzirwa zveshuga degradation (senge carbonyl compounds) zvinogona kuita nemaamino acids kuburikidza neMaillard reactions.76Kunyangwe kusviba kwemichero nemiriwo panguva yekuchengetwa kwakadzidzwa zvakanyanya, kunzwisisa kwedu maitiro aya kuchiri kushoma.77Zvichienzaniswa nenzira yeFRAP, muto weapuro wakagadziriswa laccase wakaratidzwa kushanda kwakaderera kweantioxidant nenzira yeDPPH (Tafura 2), uye kushanda kweantioxidant kwemasampuli ese kwakawedzera zvakanyanya nekuwedzera kwenguva yekuchengetedza. Nzira mbiri dzakasiyana dzekuona kushanda kweantioxidant dzakashandiswa muchidzidzo ichi nekuti misimboti yadzo yakasiyana. Nzira yeDPPH inoyera kugona kwekubvisa free radicals, nepo nzira yeFRAP ichiyera kugona kwekuderedza iron ions. Saka, zvinokurudzirwa kushandisa nzira dzakawanda dzekuona kushanda kweantioxidant kuti unzwisise zviri nani kushanda kweantioxidant yemasampuli akadzidzwa.78
Chimwe chezvakawanikwa zvikuru muchidzidzo ichi ndechekuti *Pleurotus ostreatus* laccase NRC 620 inoratidza kushanda zvakanaka pa70°C uye pH 3.0. Zvichienzaniswa nedzimwe fungus laccases dzinowanzo shandiswa pakujekesa muto, dzakadai se *Trametes versicolor* uye *Ganoderma lucidum* laccases, *P. ostreatus* NRC 620 inoratidza kugadzikana kwakanyanya kwekupisa uye pH ine acid yakawanda. Laccases kubva ku *Trametes versicolor* uye *Ganoderma lucidum* dzinowanzo ratidza kushanda kwakanaka pakati pe50-60°C uye pa pH iri pakati pe3.5 ne5.0. Musiyano uyu unogona kubatsira mukuvandudza kushanda zvakanaka kwejuice, kunyanya kune acidic juices uko kugadzikana pa pH yakaderera kwakakosha. Hunhu hwakasiyana hwe *P. Zvichienzaniswa nedzimwe fungus laccases dzakadzidzwa, *Pleurotus ostreatus* NRC 620 inoratidza kugona kushanda zvakanaka mumamiriro ezvinhu akaoma. Kupisa kwayo kwakanyanya kwekushanda zvakanaka kunoratidza zvakanakira zvinogona kuitika mumaindasitiri, zvakaita sekukurumidza kuita uye kusvibiswa kwehutachiona. pH yayo yakaderera, iyo inokodzera hunhu hweasidhi hwemajusi mazhinji, inogona kubatsira mukuita kwekujekesa muto. Izvi zvinotsigira kutsvaga kwakawanda kwekushandiswa kwakakura, zvichiita kuti *Pleurotus ostreatus* NRC 620 ive imwe nzira inoshanda pane fungus laccase sources yechinyakare. Zvichienzaniswa nezvidzidzo zvekare, takaona kuti tembiricha yepamusoro i60°C uye pH yepamusoro i3.0. Mushure mekuita pa60°C kwemaminitsi makumi masere, *Ganoderma lucidum* laccase yakachengetwa.46% yemabasa ayo.79 Sekureva kwaKurniawati naNicelle80, *Ganoderma lucidum* ma enzymes anoratidza kugadzikana kwakanaka kusvika pakati nepakati pa25°C uye pH values kubva pa5.0 kusvika 8.0, uye kugadzikana papH 6.0 uye tembiricha kubva pa10 kusvika 30°C. Muchidzidzo ichi, takaona kuti pH yakanakisa uye tembiricha yekushanda kwe enzyme ye *Pleurotus ostreatus* yaive 3.0 uye 70°C, zvichiteerana. Mushure mekubatidzwa pa40°C uye 50°C kwemaawa maviri, enzyme iyi yakachengeta 68.33% uye 59.61% yekushanda kwayo, zvichiteerana. Uyezve, Pleurotus ostreatus NRC 620 laccase yakaratidza kushanda kwakanyanya pamusoro pekupisa kwakakura kubva pa50°C kusvika 80°C, ichida kusvika pakushanda kwakanyanya (69%–98%), uye kushanda kwakanyanya kwakaonekwa pa70°C.
Mukupedzisa, howa hwe oyster laccase NRC620, hwakawanikwa pasi pemamiriro ezvinhu asingachinji, hwakaratidza kushanda zvakanaka uye kugadzikana mukati memhando dzakasiyana dze pH nemamiriro ekupisa, zvichiratidza kugadzikana kwakanyanya zvichienzaniswa nedzimwe nzvimbo dze enzyme. Kuwedzerwa kwe 10 mM MgSO₄ neCuSO₄ kwakawedzera kushanda kwe enzyme neinenge 21% ne35%, zvichiteerana. Pakagadziriswa kuita muto weapuro, enzyme yakaderedza pH uye viscosity, nepo huwandu hwe phenolic hwakaderera zvishoma panguva yekuchengetwa.
Zvakawanikwa zvinoratidza kugona kwe laccase muindasitiri yezvikafu, kunyanya mukujekesa zvinwiwa. Nekunyatso patsanura ma phenolic compounds, laccase haingoderedzi kusviba chete uye inovandudza kujeka asiwo inochengetedza mhando yemichero mumamiriro ekushanda akapfava. Kusiyana nezvinhu zvechinyakare zvekujekesa zvakaita segelatin, bentonite, uye silica gel, laccase haiburitse marara kana kubvisa hwema hunofadza kubva muzvinwiwa, zvichiita kuti ive sarudzo ine hushamwari kune nharaunda uye inogara kwenguva refu. Uyezve, kana ichienzaniswa nedzimwe enzymes uye nzira dzekusefa, laccase inopa mhinduro yakanangwa uye inodhura pasina kukanganisa kunaka kwechigadzirwa.
Kyomuhimbo, HD naBrink, HG. Mashandisirwo uye nzira dzekudzivirira kufamba kwe laccases dzine mhangura; ongororo. Heliyon 9, e13156 (2023).
Nguva yekutumira: Zvita-15-2025