Bayan kwanaki 25 na lokacin da aka saka a yanayin zafi mai tsauri a 28°C, laccase daga *Pleurotus ostreatus* NRC620 ya nuna mafi girman aiki a cikin yanayin al'adar fungal. Mafi kyawun pH da ƙimar zafin jiki na wannan enzyme sune 3.0 da 70°C, bi da bi. Bayan awanni 2 na lokacin da aka saka a 40°C da 50°C, aikin enzyme ya riƙe 68.33% da 59.61%, bi da bi. Bayan awanni 2 na lokacin da aka saka a cikin citrate-phosphate buffer (pH 7.0), aikin enzyme ya ci gaba da kasancewa a 100%. Ƙarin 10 mM MgSO₄ da CuSO₄ sun ƙara aikin enzyme da kusan 21% da 35%, bi da bi, yayin da NaCl, MnCl₂, KCl, da CaCl₂ suka hana aikin enzyme. Ta amfani da ABTS a matsayin substrate, sigogin motsi (Km da Vmax) na *Pleurotus ostreatus* NRC 620 laccase sun kasance 1.99 mM da 16,217 μmol min−1 L−1, bi da bi. Maganin enzymatic na samfuran ruwan apple ya rage pH da danko sosai, kuma wannan raguwar ya danganta da ƙaruwar lokacin ajiya. Maganin Laccase ya haifar da ɗan raguwa a cikin jimlar abubuwan phenolic na ruwan apple, amma ba a lura da raguwar ayyukan antioxidant ba.
A cikin 'yan shekarun nan, masu bincike sun mayar da hankali kan amfani da fasahar kere-kere ta halittu a masana'antar abinci. Laccase yana ɗaya daga cikin enzymes masu amfani a masana'antar abinci, yana neman aikace-aikace a fannoni kamar sarrafa ruwan 'ya'yan itace, yin burodi, daidaita ruwan inabi, da kuma inganta halayen organoleptic na kayayyakin abinci.1Yawancin tsire-tsire masu girma da ƙananan halittu suna fitar da laccase,2kuma fungi kamar deuteromycetes, ascomycetes, da basidiomycetes suma suna iya samar da laccase.3Laccase (EC 1.10.3.2) wani abu ne mai launin shuɗi wanda ke rage iskar oxygen zuwa ruwa ta amfani da tsarin da ya ƙunshi atom guda uku na jan ƙarfe, ta haka yana lalata mahaɗan phenolic daban-daban da amines masu ƙanshi. A lokacin samar da ruwan 'ya'yan itace da kayan lambu, launin ruwan kasa mai kama da na enzymatic da wanda ba enzymatic ba matsaloli ne masu mahimmanci.4Tunda waɗannan abubuwa suna yin mummunan tasiri ga launi, dandano, da ƙamshin ruwan 'ya'yan itace, dole ne a cire su.5
Daga cikin dukkan 'ya'yan itatuwa, apples sune aka fi cinyewa a duniya da kuma a Tarayyar Turai. A shekarar 2019, samar da apples ya zo na uku a duniya, inda ya zarce tan miliyan 87.6Tuffa tana ɗauke da sinadarai masu yawa na phenolic, waɗanda suka haɗa da flavonoids da phenolic acid kamar caffeic acid da chlorogenic acid.7Saboda ruwan apple yawanci ana shansa a cikin siffarsa mai haske, kusan kashi 50% zuwa 90% na abubuwan phenolic suna ɓacewa yayin aikin tacewa.8A yau, masu amfani da kayan lambu suna zaɓar samfuran da ba a sarrafa su sosai ba, kamar ruwan apple mai duhu mai yawan polyphenol. Duk da haka, saboda yawan sinadarin phenolic da ke cikinsa, wannan nau'in ruwan apple yana da saurin canzawa da duhu.9Ana amfani da fasahohi daban-daban, ciki har da hanyoyin magance zafi kamar su pasteurization a zafin 60-90°C, don rage ko hana duhun ruwan apple.10Koyaya, bisa ga binciken Sauceda-Gálvez11, sarrafa zafi na iya lalata sinadarai masu canzawa kuma yana shafar halayen organoleptic na ruwan apple. Madadin hanyoyin sarrafa zafi sun haɗa da supercritical carbon dioxide, ultraviolet radiation, ultrasound, high hydrostatic pressure, ko high-pressure homogenization.12Ingancin waɗannan fasahohin da kuma yawan ruwan 'ya'yan itace masu dacewa ya dogara ne akan ma'aunin da ake amfani da su da kuma halayen samfurin. Amfani da su yaɗuwa yana da iyaka saboda tsadar farashi, mummunan tasiri ga ingancin wasu kayayyakin abinci, ko rashin isasshen aikin enzymes.13,14
Ana iya amfani da Laccase don daidaita da kuma bayyana ruwan 'ya'yan itace.15Gökmen da sauransu.16Ana ba da shawarar amfani da laccase don fayyace ruwan 'ya'yan itace domin yana kawar da sinadarai masu guba ta hanyar mayar da su polymers ko oligomers waɗanda kowace membrane mai tacewa za a iya cire su cikin sauƙi, wanda hakan ke ba ruwan apple damar kiyaye launi da haske mai kyau har zuwa makonni shida a zafin jiki na 50°C. An sanya laccase *Trichoderma* mai tsabta a kan beads na alumina kuma an yi amfani da shi don cire sinadarai masu guba waɗanda gurɓataccen ruwan apple ya haifar.17
Kimanin kashi 80-90% na abubuwan da ke cikin ruwan apple masu canzawa sune esters da aldehydes, waɗanda ke ba da ƙamshi na musamman ga ruwan.18An cire Laccase daga *Trametes versicolor* a kan wani tallafi mai araha da aka yi da zare na halitta daga ƙananan harsashin kwakwa don tsaftace ruwan apple.19Nazarin da aka yi a baya sun binciki daidaita ruwan apple (launi da turbidity) ta amfani da hanyoyin da ba su da enzymes ko hana motsi, ko kuma tare da haɗakar tacewa ta ultrafiltration.5,19Duk da haka, tasirin laccocin fungal akan halayen sinadaran sinadaran ruwan apple yayin ajiya har yanzu ba a fayyace shi ba. Saboda haka, manufar wannan binciken ita ce a binciki canje-canjen da aka samu a cikin halayen sinadaran sinadaran, abubuwan da ke cikin mahallin phenolic, da kuma aikin antioxidant na ruwan apple bayan an yi musu magani da laccocin fungal da kuma ajiyar sati biyu a cikin firiji. Laccocin suna da ikon oxidize mahaɗan phenolic, wanda hakan ke sa su zama masu alƙawarin amfani da su a cikin hanyoyin masana'antu daban-daban, gami da bayyana ruwan 'ya'yan itace. Wannan binciken ya binciki laccocin daga *Pleurotus ostreatus* NRC 620, yana mai da hankali kan yanayin da ya dace don ayyukansu da ingancinsu wajen bayyana ruwan 'ya'yan itace. Duk da cewa bincike kan namomin kaza na kawa (P. ostreatus NRC 620) har yanzu yana da iyaka, binciken da ya gabata ya binciki enzymes daga tushen fungal daban-daban, kamar Trametes versicolor da Ganoderma lucidum. Manufar wannan binciken ita ce a tantance yuwuwar amfani da wannan enzyme a masana'antar abinci da kuma haskaka halayensa na musamman, musamman pH da zafin jiki da ya dace.
An sayi 2,2′-Azooxybis (3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) daga Sigma-Aldrich (Kanada). Duk sauran sinadaran suna da inganci na nazari.
Cibiyar Tarin Al'adun Kwayoyin Halitta ta Cibiyar Bincike ta Ƙasa ta sami sanannen nau'in namomin kaza na kawa NRC620. Bayan an yi noma a ƙarƙashin ƙasa, an adana wannan nau'in a kan ƙwanƙolin dankalin turawa na dextrose a zafin digiri 4. Hanyar shirya inoculum ita ce kamar haka: An yi wa mycelium mai shekaru 10 allurar riga-kafi a kan faranti na dankalin turawa na dextrose agar sannan aka sanya shi a zafin digiri 28. Bayan kwana 10, an cire tubalan mycelium guda uku masu diamita 12 mm daga cikin agar media ta amfani da ƙarfe mai tsafta sannan aka sanya su a cikin kwalban Erlenmeyer mai nauyin milimita 250 tare da matosai na auduga waɗanda ke ɗauke da 50 ml na maganin gargajiya mai tsafta (pH 5.0, kamar yadda Othman et al suka bayyana a baya.20). An saka al'adun a zafin digiri 28 na Celsius na tsawon kwanaki 18. Sannan aka tace al'adun ta hanyar takardar tacewa ta Whatman No. 1, kuma abin da ya fito daga ciki ya zama tushen enzyme.
An tantance aikin Laccase ta amfani da ABTS a matsayin substrate. Haɗin amsawar (2 mL) ya ƙunshi 500 μL na 0.3 mM ABTS (wanda aka narkar a cikin 0.1 M sodium citrate buffer, pH 4.5) da kuma adadin da ake buƙata na samfurin enzyme da aka narkar da shi da ruwan da aka tace.21,22Idan aka yi la'akari da cewa laccase na iya oxidize ABTS a zafin ɗaki (28 °C ± 2), an ƙaddara oxidation na ABTS ta hanyar auna ƙaruwar sha a 420 nm (ε)420= 36,000 cm-1 M -1) ta amfani da na'urar auna haske ta Agilent Carry-100 UV. An buƙaci na'urar auna haske guda ɗaya don oxidize 1 μmol ABTS a minti ɗaya. An ƙayyade yawan furotin ta hanyar hanyar Bradford ta amfani da albumin na jini na shanu a matsayin iko na ciki.23,24
Bayan samun enzyme daga nau'in naman kaza na kawa NRC 620, an auna aikinsa a tazara daban-daban na noma na tsawon kwanaki 25 a ƙarƙashin yanayin da ke canzawa a digiri 28 na Celsius.
Domin nazarin tasirin zafin jiki akan aikin laccase, an gudanar da gwaje-gwaje a cikin kewayon zafin jiki daga 20 zuwa 90 °C. Kafin ƙara enzyme da fara amsawar, an gauraya buffer (0.1 M sodium citrate, pH 4.5) da substrate (ABTS) kuma an saka su a cikin injin na tsawon mintuna 5 a yanayin zafi daban-daban. An tantance daidaiton zafin enzyme ta hanyar saka su a cikin 0.05 M sodium phosphate buffer (pH 7.0) a 40, 50, 60, da 70 °C na tsawon awanni 2, bi da bi. Sannan an tantance sauran ayyukan ta amfani da substrate ABTS.
An tantance tasirin pH akan aikin laccase ta amfani da ABTS a matsayin substrate a cikin 0.1 M citrate-phosphate buffers tare da kewayon pH na 2.5 zuwa 7.0. An saka maganin enzyme a 40°C na tsawon awanni biyu a cikin 0.1 M citrate da Tris buffers (pH 3, 4, 6, da 7) don tantance daidaiton pH. An ƙididdige sauran ayyukan tare da ABTS a matsayin substrate bayan incubation.
An saka laccase na tsawon minti 10 a cikin sinadarin sodium phosphate buffer (0.05 M, pH 7.0) wanda ke ɗauke da ions na ƙarfe daban-daban (Mg2+, Cu2+, Co2+, Ca2+, Zn2+, K+, Na+, da Mn2+) a yawan 2.5 mM da 10 mM, bi da bi. Daga nan aka ƙara substrate (ABTS) don fara amsawar, kuma aka tantance aikin da ya dace.
An auna iskar shaka ta ABTS ta hanyar laccase a wurare daban-daban (0.025–3 mM) a pH 4.5 don tantance sigogin motsi (Vmax da Km).masu daidaituwaAn ƙididdige lissafin Michaelis-Menten ta amfani da makircin Lineweaver-Burk, wanda ke nuna ma'aunin amsawar a matsayin aikin yawan abubuwan da ke cikin substrate. An ƙididdige ma'aunin motsi daga makircin Lineweaver-Burk ta amfani da software na GraphPad Prism sigar 6.01.
Bayan an wanke apples ɗin sosai da ruwan famfo, an yanke su biyu sannan aka jika su da ruwan 'ya'yan itacen apple Braun MP80 mai cikakken atomatik (wanda aka yi a Jamus). An tace ruwan 'ya'yan itacen ta hanyar mayafin cheesecloth guda huɗu. Ba a ƙara enzymes a cikin rukunin sarrafawa ba, yayin da aka ƙara 2.0% laccase (mafi inganci da aka gwada) a cikin ruwan apple da aka shirya sabo, wanda aka adana a zafin 4°C na tsawon makonni biyu.
An ƙayyade sinadarin acidity (TA) da pH bisa ga hanyar Boulton et.al.27An auna pH na kowane samfurin ta amfani da na'urar auna pH ta dijital (mita na JENWAY 3510 pH). An ƙididdige sinadarin acidity na Titratable (TA) bisa ga malic acid ta amfani da dabarar da ke ƙasa.
Inda V da C su ne girman (mL) da kuma yawan sinadarin sodium hydroxide da aka yi amfani da shi a cikin titration, bi da bi. K shine ma'aunin juyawar malic acid, daidai yake da 0.067, kuma W shine ma'aunin (g) na ruwan apple.
Jimillar daskararru masu narkewa (TDS) an tantance adadin duk samfuran ruwan 'ya'yan itace ta amfani da na'urar auna aljihu ta PAL-1 (ATAGO, Tokyo, Japan). Bayan kowace aunawa, an wanke ruwan tabarau na gani da ruwan da aka cire daga jiki, kuma an gwada kowace samfurin ruwan 'ya'yan apple sau uku. An ƙididdige ƙimar kowane samfurin ta hanyar matsakaicin ma'auni uku. An kuma ƙididdige matsakaicin karkacewar ± na kowane samfurin ruwan 'ya'yan apple ta hanyar matsakaicin waɗannan sakamakon.
An tantance ƙarfin ruwan apple ta amfani da na'urar auna juyawa (RV, Rheotest 2, Jamus). An sanya samfurin a cikin silinda "S2" na viscometer. An nuna ɗankowa ta hanyar gangaren matsin lamba na yanke da lanƙwasa ƙimar yanke, wanda aka ƙididdige shi daga matsin lamba na yanke da lanƙwasa masu dacewa a cikin ƙimar yanke daban-daban (daga 1.00 zuwa 437.4 s⁻¹). Tsarin lissafin ɗankowa mai bayyana kamar haka:
Inda η shine ɗanko da aka bayyana (cP), τ shine damuwar yankewa (dyn/cm²), γ shine ƙimar yankewa (sec⁻¹), kuma (τ) ana ƙididdige shi ta amfani da ƙimar karfin juyi (α) da silinda (Z) ta amfani da dabarar da ke ƙasa: τ = Z. α.
An ƙaddara ma'aunin launin ruwan kasa bisa ga hanyar Meidav etal.29An saka samfurin ruwan 'ya'yan itace mai nauyin 10-ml a 2750 xg na tsawon minti 10. An haɗa ruwan 'ya'yan itace mai nauyin 5 ml da 5 ml na ethanol 95%. An auna yadda ruwan ke sha a 420 nm ta amfani da na'urar auna haske ta Shimadzu UV (UV-1601 PC).
An tantance jimillar abubuwan da ke cikin phenolic (TPC) ta hanyar launi ta amfani da Folin-Ciocalteu reagent kamar yadda Boulton et al suka bayyana.[27]An gina madaidaicin layi na gallic acid don tattarawa daga 0 zuwa 500 mg/L (r²= 0.997). Sakamakon ana bayyana shi azaman daidai gwargwado na gallic acid (mg GAE/mL).
Sai a zuba 125 μL na ruwan da aka tace da kuma 2850 μL na maganin FRAP a cikin 25 μL na ruwan apple sannan a bar hadin ya yi duhu don ya yi kauri.30Minti. Sannan a auna yadda ake sha a 593 nm ta amfani da na'urar auna haske ta Shimadzu UV (UV-1601 PC). An shirya sinadarin FRAP ta hanyar haɗa sinadarin acetate buffer 300 mM (pH 3.6), ƙarfe 20 mM (III) chloride, da 10 mM 2,4,6-tris(2-pyridyl)triazine (TPTZ) (wanda aka narkar a cikin 40 mM HCl) a cikin rabo na 10:1:1. An samar da lanƙwasa ta yau da kullun ta amfani da Trolox a matsayin ma'auni (R²= 0.999), kuma an bayyana sakamakon a matsayin μM Trolox/mL.
An tantance tasirin maganin hana tsufa na ruwan 'ya'yan itace da aka yi wa magani da kuma wanda ba a yi wa magani ba ta hanyar amfani da hanyar DPPH don tantance ikonsu na korar radicals masu kyauta na DPPH.31An gauraya ƙananan lita goma na ruwan 'ya'yan itace da 1 ml na maganin DPPH (100 μM) a cikin methanol. Bayan amsawa a cikin duhu na tsawon mintuna 30, an auna shan ruwan cakuda a 517 nm ta amfani da na'urar auna haske ta Shimadzu UV (UV-1601 PC). An bayyana sakamakon a matsayin daidai gwargwado na trolox (μM trolox/ml) bisa lanƙwasa na daidaitawa (R2= 0.990).
Bayanan da aka samu sun nuna cewa an lura da yawan samar da laccase a cikin namomin kaza na NRC 620 a ƙarshen rana ta 18 ta fermentation, inda suka kai ga aikin 1302 U/L. Wannan ya zama tushen tantance lokacin noma mafi kyau don samar da laccase (Hoto na 1). Duk da cewa samar da enzyme ya ƙaru tare da ƙaruwar lokacin noma, ƙimar ƙaruwar ba ta yi daidai da lokacin noma ba; bayan kwanaki 21, aikin enzyme ya ƙaru da 90 U/L kawai (zuwa 1390 U/L). Saboda haka, a ƙarshe an zaɓi kwanaki 18 a matsayin lokacin noma mafi kyau don daidaita yawan amfanin gona tare da fa'idodin tattalin arziki na ƙaruwar lokacin noma.
Tasirin lokacin noma akan yawan amfanin gona na laccase a cikin Pleurotus ostreatus NRC 620. An yi allurar tubalan mycelium guda uku (12 mm) a cikin 50 ml na maganin da ba shi da tsafta sannan aka dasa su a zafin jiki na 28 °C na lokuta daban-daban.
Kamar yadda aka yi a wasu bincike, sakamakonmu ya nuna cewa lokacin da ya dace don samun kololuwar fitar da laccase daga fungi yana iya kasancewa tsakanin kwanaki 7 zuwa 36.32A cewar Ezike da sauransu.33, *Trametes polyzona* WRF03 ya samar da mafi girman adadin laccase a ƙarshen rana ta tara ta fermentation, tare da takamaiman aikin furotin 1637 U/mg. Bugu da ƙari, Othman et al.34sun gano cewa *Trichoderma harzianum* S7113 ya fitar da adadi mai yawa na laccase a rana ta biyar ta al'ada. Yawan samar da laccase ya kai kololuwar aiki a rana ta goma sha huɗu sannan a hankali ya ragu.34Duk da cewa fitar da enzyme na iya faruwa a lokacin babban matakin girma, yawanci yakan kai kololuwa a lokacin matsakaicin lokaci kuma yana faruwa ne ta hanyar amfani da tushen carbon ko nitrogen.34,35
Duk da cewa laccase daga Pleurotus ostreatus NRC 620 ya nuna babban aiki a cikin kewayon zafin jiki mai faɗi daga 50°C zuwa 80°C, yana gab da kusantar aiki mafi girma (69–98%), an lura da matsakaicin aikin sa a 70°C (Hoto na 2a). A wajen wannan kewayon zafin jiki, aikin enzyme ya ragu a kusan 70°C. Waɗannan sakamakon sun nuna cewa enzyme yana aiki a yanayin zafi mai yawa, wataƙila saboda yawan zafin jiki yana ƙara kuzarin motsi na amsawar.
Tasirin zafin amsawa (a) da pH (b) akan aikin laccase a cikin *Pleurotus ostreatus* NRC 620. An cimma yanayin zafi tsakanin 20 zuwa 90 °C ta hanyar sanya cakuda a cikin yanayin zafi daban-daban na tsawon mintuna 5 kafin a ƙara enzyme da kuma fara amsawar. An tantance tasirin pH akan aikin laccase ta amfani da ABTS a matsayin substrate a cikin mafita waɗanda ke ɗauke da 0.1 M citrate-phosphate buffer akan kewayon pH na 2.5 zuwa 7.0.
A cewar Ezike etal.33, mafi kyawun zafin jiki don *Trametes polyzona* WRF03 laccase shine 55 °C, wanda yayi daidai da na *Ganoderma lucidum*laccase36kuma yayi kama da mafi kyawun zafin jiki (50 °C) don *Trametes polyzona* KU-RNW02737laccase . Baldrian38ya lura cewa, kamar sauran tsarin enzyme masu lalata lignin, yanayin zafin da ya dace don laccase yana tsakanin 50 zuwa 70 °C.
Sakamakon ya nuna cewa enzyme ɗin ya nuna mafi girman aiki a pH 3.0, inda ya kai kashi 94% na aiki a pH 3.5. Duk da haka, ya ci gaba da aiki a kan kewayon pH mai faɗi daga 2.5 zuwa 7.0 (Hoto na 2b). Bugu da ƙari, ya nuna mafi girman aiki a cikin yanayin acidic idan aka kwatanta da yanayin tsaka-tsaki ko alkaline. Ayyukansa sun kasance aƙalla kashi 77% akan kewayon pH daga 2.5 zuwa 4.5, amma sun kai kusan kashi 38% kawai a pH 7.0. Mafi kyawun pH don laccase daga *Trametes polyzona* WRF03 shine 4.533, wanda yayi daidai da pH na laccase daga *Trametes polyzona* KU-RNW02737, *Trichoderma harzanium* 39, *Pleurotus* sp. 40, da *Trametes hirsuta* 41. Duk da haka, a cewar binciken da Chairin et al.42, mafi kyawun pH don laccase daga *Polymorpha f. sp.* WR710-1 shine 2.2, yayin da mafi kyawun pH don laccase daga *Polymorpha f. sp.* IBL-04 shine 5.043. Haɗa anions na hydroxide (laccase inhibitor) zuwa atom ɗin jan ƙarfe na laccase na T2/T3 na iya zama dalilin raguwar aikin laccase a ƙarƙashin yanayin pH tsaka tsaki ko alkaline. Wannan na iya kawo cikas ga canja wurin electron na ciki daga cibiyar T1 zuwa cibiyar T2/T3, don hakaiyakanceaikin enzyme23,44
Ta hanyar sanya enzyme a yanayin zafi daban-daban, an gano cewa lokacin shiryawa da zafin jiki sun shafi daidaiton enzyme. Abin lura shi ne, laccase daga *Trametes polyzona* NRC 620 ya nuna kwanciyar hankali mafi girma a 40℃ da 50℃, yana riƙe da 68.33% da 59.61% na aikin farko, bi da bi, bayan mintuna 120 (Hoto na 3a). Sabanin haka, a ƙarƙashin yanayi iri ɗaya (40℃ da 50℃, mintuna 120), laccase daga *Trametes polyzona* WRF03 ya riƙe 64.38% da 42.92% na aikinsa, bi da bi.33Akasin haka, ƙara lokacin ƙunƙuwa da zafin jiki sun rage kwanciyar hankali na *Trametes polyzona* NRC 620 laccase; Bayan ƙunƙuwa a 60℃ da 70℃ na tsawon mintuna 60, aikinsa ya ragu zuwa 39.24% da 1.72%, bi da bi (Hoto na 3a). Daidai da sakamakon gwaji, laccase daga *Trametes polyzona* WRF03 ya nuna kwanciyar hankali mafi girma a 40 da 50℃ a duk tsawon aikin maganin zafi.33Hakazalika, Lueangjaroenkit etal.37da kuma Chairin etal.42ya ba da rahoton daidaiton laccases daga Trametes polyzona KURNW027 da Trametes polyzona WR710-1 a zafin 50 °C na tsawon awa 1, bi da bi. A matsayin mai amfani da biocatalyst da ke aiki a fannoni daban-daban na fasahar halittu, laccase ya kamata ya sami kyakkyawan kwanciyar hankali da aiki a kan kewayon zafin jiki mai faɗi.
Kwanciyar thermostatic (a) da kwanciyar pH (b) na laccase daga *Pleurotus ostreatus* NRC 620. An tantance kwanciyar hankali na thermostatic ta hanyar sanya maganin enzyme a cikin sodium phosphate buffer mai nauyin 0.05 M (pH 7.0) a digiri 40, 50, 60, da 70 °C na tsawon awanni 2, bi da bi. An tantance kwanciyar hankali na pH ta hanyar sanya maganin enzyme a cikin 0.1 M citrate buffer da Tris buffer (pH 3, 4, 6, da 7) a digiri 40 °C na tsawon awanni 2. An ƙididdige sauran ayyukan ta amfani da ABTS a matsayin substrate bayan kunciyar.
Domin tantance yanayin da ya dace don amfani da adana enzyme, mun binciki tasirin pH akan kwanciyar hankali na laccase. Fuskantar nau'ikan pH daban-daban ya shafi kwanciyar hankali na tsarin furotin, wanda hakan ya shafi kwanciyar hankali da aikin kwayar halittar enzyme. Sakamakon ya nuna cewa enzyme bai da kwanciyar hankali a ƙarƙashin yanayin acidic, yayin da ya nuna ingantaccen kwanciyar hankali a mafi girman ƙimar pH (yankunan tsaka tsaki da alkaline). A ƙimar pH na 7.0, 6.0, 4.0, da 3.0, ƙimar riƙe enzyme bayan mintuna 120 sun kasance kusan 100%, 62.54%, 52.39%, da 11.14%, bi da bi (Hoto na 3b). *Strombus multisus* Laccase WRF03 ya nuna kwanciyar hankali mafi girma a ƙimar pH tsaka tsaki (5.5-6.5) da ƙarancin kwanciyar hankali a ƙimar pH mai acidic (ƙasa da 4.0). Bayan mintuna 120 a ƙimar pH na 5.5, 6.0, da 6.5, ƙimar riƙewar enzyme ta kasance kusan kashi 82%, 100%, da 93%, bi da bi.33Khairin da sauransu.42ya lura cewa laccase daga Trametes polyzona WR710-1 ya kasance mai karko a cikin kewayon pH na 6.0 zuwa 7.0, yayin da Sayed et al.45ya nuna cewa laccase ya fi karko a ƙarƙashin yanayin pH mai tsaka-tsaki. Duk da haka, laccase daga Cerrena unicolor shi ma ya nuna karko a ƙarƙashin yanayin alkaline (pH 9.0)46Laccases ɗin da aka yi nazari a kansu sun nuna kwanciyar hankali mai yawa a kan kewayon pH mai faɗi. Wannan na iya zama muhimmin siffa ga aikace-aikacen masana'antu.
Tunda wasu ions na ƙarfe suna da tasirin ƙarfafawa da hana aiki akan ayyukan enzyme, dole ne a yi la'akari da tasirinsu akan ayyukan enzyme a aikace-aikacen masana'antu. Wannan yana da mahimmanci saboda ions na ƙarfe gurɓatattun abubuwa ne na muhalli waɗanda zasu iya shafar kwanciyar hankali da haɗa enzymes na waje.47Domin bincika tasirin ions na ƙarfe da yawa akan laccase daga *Pleurotus ostreatus* NRC 620, mun gudanar da gwaje-gwaje masu dacewa. Kamar yadda aka nuna a Hoto na 4, ya danganta da nau'in ƙarfe da aka yi amfani da shi, ƙara yawan ions na ƙarfe daga 2.5 mM zuwa 10 mM ya shafi aikin enzyme. Misali,Mg²⁺ , Co²⁺ , Zn²⁺, kumaCu²⁺yana iya ƙarfafa aikin enzymes da kuma kunna su,Na⁺ , Mn²⁺ , Ca²⁺, kumaK⁺zai iya hana aikin enzyme. A yawan 10 mM, ions na Cu²⁺ da Mg²⁺ sune mafi ƙarfin kunnawa na aikin laccase daga *Pleurotus ostreatus* NRC 620, suna samar da matakin kunnawa na kusan 34% da 20%, bi da bi. Duk da haka, a yawan 10 mM, ions na Ca²⁺ sune mafi ƙarfin hana laccase, suna rage aikin enzyme da kusan 60%.
Tasirin ions na ƙarfe akan aikin Pleurotus ostreatus NRC 620 laccase. An saka laccase na tsawon mintuna 10 a cikin sinadarin sodium phosphate buffer (0.05 M, pH 7.0) wanda ke ɗauke da ions na ƙarfe daban-daban a cikin yawan 2.5 mM da 10 mM. Daga nan aka fara amsawar ta hanyar ƙara substrate (ABTS), bayan haka aka auna aikin da ya dace.
Sakamakonmu ya yi daidai da na wasu marubuta waɗanda suka gano cewa Mg²⁺ da Cu²⁺ suna haɓaka aikin *Trametes polyzona* WRF03³. Castaño et al.⁴⁸ sun gano cewa laccase daga *Xylaria* sp. ana motsa shi zuwa wani mataki ta hanyar ions na jan ƙarfe (Cu²⁺). Bugu da ƙari, Foroutanfar et al.⁴⁹ da Si et al.⁵⁰ sun gudanar da irin wannan bincike kan laccase daga *Paraconiothyrium variabile* da *Trametes pubescens*, bi da bi. Wurin ɗaure jan ƙarfe na nau'in II (T2) na wannan enzyme za a iya cika shi da Cu²⁺ a wani taro da aka bayar, wanda zai iya bayyana yadda aikin laccase ke ƙaruwa a mafi yawan Cu²⁺³⁹. Tunda laccases na fungi masu launin fari suna ɗauke da ƙwayoyin jan ƙarfe da yawa, tasirin ions na jan ƙarfe akan aikin laccase ya bambanta kuma ya kama daga mai motsa jiki da hanawa zuwa tsaka tsaki.⁵¹ Sabanin haka, Zhou et al.. [52]ya ruwaito cewaCu²⁺ya hana aikin laccase na termite na ƙarƙashin ƙasa na Taiwan (Odontotermes formosanus). Duk da haka, laccases na Cerena sp. HYB07[53]da kuma Clitocybe maxima[54]ba su shafi ions na jan ƙarfe ba.
An wakilta takamaiman yanayin substrate ta hanyar sigogin motsi (Km da Vmax); ƙarfin haɗin da substrate ke da shi da enzyme, ƙarancin ƙimar Km kuma mafi girman takamaiman yanayin substrate.3,21,55An tantance sigogin motsi (Km da Vmax) na laccase daga *Pleurotus ostreatus* NRC 620 ta amfani da software na GraphPad Prism 6.0 ta hanyar zana taswirar Lineweaver-Burk (Hoto na 5). Lokacin amfani da ABTS azaman substrate, sakamakon shine 1.99 mM da 16217 μmol.minti⁻¹ L⁻¹,bi da bi. Elsayed da sauransu.21ya ruwaito cewa ƙimar Km don oxidation na ABTS sun kasance 0.1 mM da 0.064 mM, bi da bi, yana nuna babban alaƙar isoenzymes na Lac A da Lac B ga ABTS. Bugu da ƙari, ƙimar Vmax sun kasance 0.182 μmolminti⁻¹da kuma 0.603 μmolminti⁻¹, bi da bi. Ƙimar Km da aka samu ta yi ƙasa da ta Trametes polyzona WRF03 (8.66 mM); haka kuma, ƙimar Vmax ɗinsu (1429 mmol min⁻¹) ita ma ta kasanceƙasalokacin amfani da ABTS a matsayin substrate.33 Hakazalika, ƙimar Km na Lentinus squarrosulus MR13 da Trametes sp. AH28-2 laccase yawansu ya kasance 0.0714 mM da 0.025 mM, bi da bi, kuma ƙimar Vmax sun kasance 0.0091 mM min−1 da 0.67 mM min−1 mg−1 (idan aka kwatanta da ABTS), bi da bi.56,57
An binciki tasirin yawan ABTS akan aikin laccase daga *Pleurotus ostreatus* NRC 620, kuma an zana taswirar Lineweaver-Burk na musayar saurin amsawar farko idan aka kwatanta da yawan ABTS. An auna amsawar oxidation na ABTS tare da yawan (0.025–3.0 mM) na laccase a pH 4.5 don tantance sigogin motsi (Vmax da Km). An ƙididdige ma'aunin motsi na Michaelis-Menten ta amfani da taswirar Lineweaver-Burk na musayar saurin amsawa idan aka kwatanta da yawan substrate. An ƙididdige ma'aunin motsi daga taswirar Lineweaver-Burk ta amfani da software na GraphPad Prism 6.01.
Enzymes na gargajiya masu bayyanawa, kamar pectinases, hydrolyze pectic acid, rage danko da turbidity. Suna wargaza polysaccharides na tsari yadda ya kamata kuma galibi ana amfani da su tare da wasu enzymes, kamar cellulases da hemicellulases, don inganta yawan amfanin ƙasa da haske. Duk da haka, pectinases ba sa kai hari ga mahaɗan phenolic musamman, waɗanda sune manyan abubuwan da ke ba da gudummawa ga turbidity da oxidative browning, musamman a cikin ruwan 'ya'yan itace kamar apple da ruwan inabi.58Sabanin haka, laccases suna haɓaka iskar shaka ta mahaɗan phenolic, suna mai da su manyan ƙwayoyin halitta marasa narkewa waɗanda za a iya cire su ta hanyar narkewa ko tacewa. Wannan hanyar ba wai kawai tana inganta haske ba ne, har ma tana ƙara tsawon rayuwar ruwan 'ya'yan itace ta hanyar rage yuwuwar launin ruwan kasa mai kauri wanda mahaɗan phenolic ke haifarwa. Bugu da ƙari, ana iya aiwatar da hanyoyin bayyanawa bisa laccase a ƙarƙashin yanayi mai sauƙi na sarrafawa (pH 3.5–5.5, zafin jiki 25–40 °C), wanda hakan ke sa su dace da ruwan 'ya'yan itace masu laushi ba tare da lalata halayen abinci mai gina jiki ko na organoleptic ba.59Bincike ya nuna cewa maganin pectinase zai iya tsarkake ruwan 'ya'yan itace cikin awanni 1-2, yayin da maganin laccase yawanci yana buƙatar lokaci mai tsawo (awanni 3-6) don rage mahaɗan phenolic gaba ɗaya. Duk da haka, ana iya inganta wannan tsari ta hanyar rage motsi na enzyme ko ta hanyar haɗa laccase da hanyoyin bayyanawa na inji.60A cikin wannan binciken, bayanin enzyme na ɗanyen da aka cire ya nuna manyan ayyukan laccase da α-amylase, yayin da ayyukan pectinase da xylanase suka yi ƙasa sosai, kuma ba a gano ayyukan cellulase ba. Saboda haka, raguwar turbidity da abun ciki na phenolic ya faru ne galibi saboda aikin laccase, yayin da canjin danko na iya zama wani ɓangare saboda aikin amylase.
Tebur na 1 yana nuna ma'aunin sinadaran sinadaran ruwan apple da aka matse sabo da samfuran da aka matse sabo. Sakamakon ya nuna cewa yawan ruwan apple da aka matse sabo (71.59%) ya yi ƙasa da na samfuran da aka matse sabo (87.34%). Waɗannan sakamakon sun yi daidai da binciken Pilnik da Orange.61, wanda ya nuna cewa amfani da enzymes a cikin sarrafa 'ya'yan itace na iya ƙara yawan ruwan 'ya'yan itace, inganta tacewa, da kuma samun ruwan 'ya'yan itace mai inganci da tsafta don tattarawa. Ƙara yawan ruwan 'ya'yan itace ya faru ne galibi saboda ƙaruwar yawan sukari mai narkewa a cikin ruwan. A lokacin da ake amfani da enzymes hydrolysis na 'ya'yan itatuwa, mesoglea da pectin a cikin bangon tantanin halitta na samfurin ana lalata su kuma ana canza su zuwa abubuwa masu narkewa kamar sukari da acid masu tsaka tsaki.62.Darajar pH na ruwan apple da aka yi wa magani da enzyme ya yi ƙasa sosai da na ƙungiyar kulawa (P < 0.05), kuma ƙimar pH na ƙungiyoyin biyu ta ƙaru sosai yayin ajiya (Tebur 1). Waɗannan sakamakon sun yi daidai da na Mark et al.63, wanda ya lura cewa pH na ruwan 'ya'yan itacen cashew ya ragu bayan ajiya bayan maganin zafi. Rushewar pectin da samuwar galacturonic acid bayan maganin enzyme na iya zama sanadin karuwar pH yayin ajiya. pH na samfuran da aka yi wa enzyme magani ya kasance tsakanin 4.05 da 4.31 a duk lokacin ajiya, yayin da pH na ruwan apple da ba a yi wa magani ba ya kasance tsakanin 4.12 da 4.33.
Jimlar sinadarin acid (TA) na samfuran da ba a yi musu magani ba da waɗanda aka yi musu magani da waɗanda aka yi musu magani da laccase sun nuna raguwar yanayin tare da ƙaruwar lokacin ajiya (Tebur 1). An danganta raguwar sinadarin acid ɗin ne da sauya sinadarin organic acid zuwa carbohydrates ko halayen enzymatic, da kuma iskar shaka yayin adana ruwan 'ya'yan itace.64Jimillar sinadarin acid na ruwan apple mai sarrafawa da samfuran da aka yi wa magani da enzyme ya yi ƙasa da na sauran ruwan 'ya'yan itace (ruwan strawberry 0.9%, ruwan plum 2.2%, ruwan kumquat 1.0%, ruwan apricot 2.4%, ruwan lemu 0.8%), amma yayi kama da na sauran ruwan 'ya'yan itace (misali, ruwan pear 0.3%).62Waɗannan bambance-bambancen da ke cikin ruwan apple da aka matse ba tare da an yi masa magani ba na iya faruwa ne saboda dalilai daban-daban kamar yanayin girma, abubuwan da suka shafi kwayoyin halitta, matakin girma, da hanyoyin sarrafawa.65Rage yawan sinadarin acid da ruwan apple da aka yi wa magani da laccase ya yi daidai da sakamakon da Singh da abokan aikinsa suka gabatar.66game da raguwar jimlar sinadarin acid na ruwan apple na Jin Nuo bayan kwana 74 na ajiya. A gefe guda kuma, Oshmiansky da Wojdylo67bai sami wani muhimmin canji a cikin sinadarin acid na ruwan apple ba lokacin da ake nazarin tasirin hanyoyin bayyanawa na gargajiya.
Sakamakon da aka gabatar a cikin Jadawali na 1 ya nuna cewa jimlar ƙimar ruwan apple da aka yi wa laccase magani (TSS) ya fi na samfurin da ba a yi wa magani ba. Waɗannan sakamakon sun yi daidai da binciken da aka buga.. 68Bugu da ƙari, Tebur 1 ya nuna cewa ƙimar TSS na rukunin ruwan apple mai sarrafawa shine 9.58 a farkon lokacin kuma ya kai 11.05 a ƙarshen lokacin ajiya. Waɗannan ƙimar sun yi ƙasa da ƙimar TSS na ruwan apple sabo da Hamid et al suka ruwaito.. 69(11.2 da 11.80, bi da bi). Ƙimar TSS na samfuran ruwan apple da aka yi wa laccase ya ƙaru sosai, tun daga 11.23 kuma ya kai 12.93 bayan makonni biyu na ajiya a 4°C (Tebur 1). An kuma lura da irin wannan ƙaruwa a cikin TSS yayin ajiya a cikin 'ya'yan itatuwa citrus, lemun tsami, da lemu mai zaki. Ƙara yawan daskararru masu narkewa (TSS) yayin ajiya na iya zama saboda hydrolysis na polysaccharides (sitaci) zuwa monosaccharides (sukari), ƙaruwar yawan danshi saboda bushewar ruwan 'ya'yan itace, da kuma lalacewar pectin a cikin ruwan zuwa daskararru masu narkewa. Ƙara yawan daskararru masu narkewa (TSS) wataƙila ya faru ne saboda ƙaruwar daskararru masu narkewa, wanda za a iya samu ta hanyar canza pectin ko cellulose zuwa sukari mai narkewa ta hanyar pectin ko cellulose, bi da bi, ko ta hanyar hydrolysis na sitaci zuwa sukari, kamar yadda Hamed et al. suka ruwaito.69.Ana iya ganin tasirin laccase akan halayen ruwan apple a gani, domin ruwan apple da aka yi wa laccase yana nuna kyakkyawan sauƙin kwarara da ƙarancin ɗanko fiye da ruwan apple da ba a yi wa magani ba. An rubuta wannan lura a cikin Jadawali na 1; Danko na samfurin da aka yi wa enzyme shine 1.87 cP, yayin da danko na samfurin sarrafawa shine 2.95 cP. Wannan raguwa mai mahimmanci a cikin danko yana yiwuwa ne saboda ƙarfin riƙe ruwa na abubuwa masu kama da pectin da kuma samuwar tsarin hanyar sadarwa mai haɗin kai.
A cikin wannan binciken, an binciki tasirin laccase akan ma'aunin launin ruwan kasa (BI) na ruwan apple ta hanyar auna shan ruwa a 420 nm ta amfani da na'urar auna haske (spectrophotometer). Sakamakon an nuna shi a cikin Jadawali na 1. A lokacin ajiya, BI na samfuran ruwan apple a cikin ƙungiyoyin da aka yi wa magani da waɗanda ba a yi wa magani ba sun nuna ƙaruwa a hankali. BI yana nuna matakin launin ruwan kasa kuma yana iya zama kamarwani muhimmin abualamar halayen launin ruwan kasa na enzyme da waɗanda ba enzymatic ba. Sha yana ƙaruwa sosai yayin ajiya (P < 0.05). A ƙarshen ajiya,A420Darajar samfuran ruwan apple a cikin rukunin sarrafawa da waɗanda aka yi wa magani da enzyme ya ƙaru da kusan kashi 217% da 121%, bi da bi (Tebur 1). Sakamakon ya nuna cewa maganin enzyme zai iya rage matakin launin ruwan kasa da kusan kashi 56%. Sakamakon Bezerra et al.[19]] sun yi daidai da sakamakonmu; Sun yi amfani da zare na laccase-glutaraldehyde-coconut don bayyana ruwan apple, wanda ya rage launinsa na asali da kashi 61%.
Duk da cewa polyphenols da ke cikin ruwan 'ya'yan itace suna da tasirin abinci mai gina jiki da warkarwa mai kyau ga jikin ɗan adam, suna iya amsawa da sunadaran, suna haifar da gajimare, laka, ko danshi, ta haka suna canza dandano da ƙamshin samfurin kuma suna rage tsawon lokacin da zai ɗauka.71Manufar wannan binciken ita ce a rage yawan sinadarin phenolic da ke cikin ruwan apple ta amfani da laccase daga Pleurotus ostreatus NRC 620 cikin aminci. Sakamakon da aka gabatar a cikin Jadawali na 1 ya nuna cewa jimillar sinadarin phenolic da ke cikin ruwan apple da aka yi wa laccase magani ya ragu sosai kafin a adana shi a zafin digiri 4 na Celsius. Bugu da ƙari, jimlar sinadarin phenolic shi ma ya ragu yayin ajiya a cikin samfuran biyu da aka yi nazari a kansu (Jadawali na 1). Binciken da Sandri et al.72ya nuna cewa ruwan apple da aka yi wa magani da enzyme zai iya riƙe aikinsa na hana tsufa da kuma sinadarin phenolic. Duk da haka, sakamakon wani bincike da Lettera et al.73nuna cewa maganin ruwan lemu da fungal laccase na iya rage yawan sinadarin phenolic a cikinsa da kashi 45%.
An nuna cewa mahaɗan phenolic suna da halaye kamar su cire ƙwayoyin cuta masu guba, rage iskar oxygen da kashe su, canja wurin sinadarin hydrogen atom, da kuma bayar da electrons ga ƙwayoyin cuta masu guba, wanda hakan ke sa su zama masu ƙarfi wajen hana ƙwayoyin cuta.74Saboda haka, a cikin wannan binciken, an yi amfani da hanyoyin DPPH da FRAP don tantance tasirin laccase akan aikin antioxidant na ruwan apple da aka adana a cikin firiji na tsawon kwanaki 14 (Tebur 2). Duk hanyoyin sun nuna ƙaruwar aikin antioxidant yayin ajiya, wanda zai iya zama saboda ƙaruwar mahaɗan phenolic kyauta ko samuwar samfuran amsawar Maillard (MRPs), tare da samfuran amsawar Maillard wataƙila shine sanadin ƙaruwar aikin antioxidant.75Halayen launin ruwan kasa marasa enzymatic (gami da lalacewar ascorbic acid, halayen Maillard, da lalacewar sukari da acid ke haifarwa) suna samar da launin ruwan kasa (melanoidins). Kayayyakin lalacewar ascorbic acid na matsakaici da kayayyakin lalacewar sukari (kamar mahaɗan carbonyl) na iya amsawa da amino acid ta hanyar halayen Maillard.76Duk da cewa an yi nazari sosai kan yadda ake yin launin ruwan kasa ga 'ya'yan itatuwa da kayan lambu yayin ajiya, fahimtarmu game da waɗannan halayen har yanzu tana da iyaka.77Idan aka kwatanta da hanyar FRAP, ruwan apple da aka yi wa laccase magani ya nuna ƙarancin aikin antioxidant ta hanyar hanyar DPPH (Tebur 2), kuma aikin antioxidant na duk samfuran ya ƙaru sosai tare da ƙaruwar lokacin ajiya. An yi amfani da hanyoyi guda biyu daban-daban don tantance aikin antioxidant a cikin wannan binciken saboda ƙa'idodinsu sun bambanta. Hanyar DPPH tana auna ikon kawar da free radicals, yayin da hanyar FRAP ke auna ikon rage ions na ƙarfe. Saboda haka, ana ba da shawarar amfani da hanyoyi da yawa don tantance aikin antioxidant don fahimtar aikin antioxidant na samfuran da aka yi nazari a kansu.78
Ɗaya daga cikin mahimman abubuwan da aka gano a wannan binciken shine cewa *Pleurotus ostreatus* laccase NRC 620 yana nuna aiki mafi kyau a 70°C da pH 3.0. Idan aka kwatanta da sauran laccase na fungal da aka saba amfani da su don bayyana ruwan 'ya'yan itace, kamar *Trametes versicolor* da *Ganoderma lucidum* laccase, *P. ostreatus* NRC 620 yana nuna kwanciyar hankali mafi girma na zafi da pH mai yawan acidic. Laccase daga *Trametes versicolor* da *Ganoderma lucidum* yawanci suna nuna aiki mafi kyau a cikin kewayon 50-60°C da kuma a ƙimar pH tsakanin 3.5 da 5.0. Wannan bambanci na iya taimakawa wajen inganta ingancin bayyana ruwan 'ya'yan itace, musamman ga ruwan 'ya'yan itace mai yawan acidic inda kwanciyar hankali a ƙananan ƙimar pH yake da mahimmanci. Halaye na musamman na *P. Idan aka kwatanta da sauran laccase na fungal da aka yi nazari a kansu, *Pleurotus ostreatus* NRC 620 yana nuna ikon yin aiki yadda ya kamata a ƙarƙashin yanayi mafi ƙalubale. Mafi girman zafin jiki mafi kyau na aikinsa yana nuna fa'idodi masu yuwuwa a aikace-aikacen masana'antu, kamar saurin amsawa da rage gurɓatar ƙwayoyin cuta. Ƙananan pH ɗinsa, wanda ya dace da yanayin acidic na yawancin ruwan 'ya'yan itace, na iya zama da amfani a cikin hanyoyin bayyana ruwan 'ya'yan itace. Waɗannan sakamakon sun ba da hujjar ƙarin bincike don amfani da babban sikelin, wanda ya sa *Pleurotus ostreatus* NRC 620 ya zama madadin hanyoyin laccase na fungal na gargajiya. Idan aka kwatanta da binciken da suka gabata, mun gano cewa mafi kyawun zafin jiki shine 60°C kuma mafi kyawun pH shine 3.0. Bayan amsawa a 60°C na mintuna 80, an riƙe *Ganoderma lucidum* laccase46Kashi 79 cikin ɗari na ayyukansa. A cewar Kurniawati da Nicelle80, *Ganoderma lucidum* enzymes suna nuna kyakkyawan kwanciyar hankali zuwa matsakaici a 25°C da ƙimar pH tsakanin 5.0 zuwa 8.0, da kwanciyar hankali a pH 6.0 da yanayin zafi tsakanin 10 zuwa 30°C. A cikin wannan binciken, mun gano cewa mafi kyawun pH da zafin jiki don aikin enzyme ga *Pleurotus ostreatus* sune 3.0 da 70°C, bi da bi. Bayan shiryawa a 40°C da 50°C na tsawon awanni biyu, enzyme ɗin ya riƙe 68.33% da 59.61% na aikinsa, bi da bi. Bugu da ƙari, Pleurotus ostreatus NRC 620 laccase ya nuna babban aiki a kan kewayon zafin jiki mai faɗi daga 50°C zuwa 80°C, kusan isa matsakaicin aiki (69%–98%), tare da matsakaicin aiki da aka lura a 70°C.
A ƙarshe, laccase na naman kaza na oyster NRC620, wanda aka samu a ƙarƙashin yanayin da ba ya canzawa, ya nuna ingantaccen aiki da kwanciyar hankali a cikin yanayin pH da yanayin zafin jiki, yana nuna ingantaccen kwanciyar hankali idan aka kwatanta da sauran hanyoyin enzyme. Ƙarin 10 mM MgSO₄ da CuSO₄ ya ƙara yawan aikin enzyme da kusan 21% da 35%, bi da bi. Lokacin da aka sarrafa shi zuwa ruwan apple, enzyme ya rage pH da danko, yayin da abun ciki na phenolic ya ragu kaɗan kawai yayin ajiya.
Sakamakon ya tabbatar da yuwuwar laccase a masana'antar abinci, musamman a fannin bayyana abubuwan sha. Ta hanyar rarraba sinadaran phenolic, laccase ba wai kawai yana rage datti da inganta tsabta ba, har ma yana kiyaye ingancin ruwan 'ya'yan itace a ƙarƙashin yanayi mai sauƙi na aiki. Ba kamar sinadaran bayyana abubuwa na gargajiya kamar gelatin, bentonite, da silica gel ba, laccase ba ya haifar da sharar gida ko cire ƙamshi mai daɗi daga abubuwan sha, wanda hakan ya sa ya zama zaɓi mafi dacewa ga muhalli da dorewa. Bugu da ƙari, idan aka kwatanta da sauran enzymes da hanyoyin tacewa, laccase yana ba da mafita mai kyau da araha ba tare da lalata ingancin samfur ba.
Kyomuhimbo, HD da Brink, HG. Amfani da dabarun hana motsi na laccoci masu ɗauke da jan ƙarfe; bita. Heliyon 9, e13156 (2023).
Lokacin Saƙo: Disamba-15-2025