Produkcija glivnih lignocelulolitičnih encimov na trdnem odpadku iz papirne industrije.

Jurinčič, Nadja (author), Vodovnik, Maša (mentor)

V papirniški industriji nastaja velika količina odpadnih muljev, ki predstavljajo okoljski in ekonomski problem. V magistrskem delu smo želeli dokazati, da lahko odpadnemu mulju damo dodano vrednost, s tem da bi ga uporabili kot substrat za rast gliv iz katerih bi pridobivali lignocelulolitične encime. Preizkusili smo rast 29 različnih sevov gliv na papirniškem mulju ter določili 10 sevov (H35, CHP 4, PP3, PLO 4, HYUL, POLY xy, AASC, COCO, GAL 5, PLO ZABR), ki so najbolje preraščali substrat. Analizirali smo encimske aktivnosti v ekstraktih izbranih sevov ter izbrali tri seve. H35, PP3 in HYUL smo ponovno nacepili na mulj, ki smo ga predhodno obdelali z avtoklaviranjem 1 uro pri temperaturi 124 °C in tlaku 1,3 bar, toplotno obdelali pri 60° C ter na toplotno neobdelani mulj. Ugotovili smo, da glive na mulju, ki je bil toplotno obdelan pri 60° C ne rastejo ter da na avtoklaviranem in neobdelanem mulju najbolje uspeva sev HYUL. Slednjega smo v različnih fazah rasti homogenizirali in pripravili grob encimski ekstrakt v katerem smo določali celulolitično, ksilanolitično in lakazno aktivnost. Aktivnost hidrolitičnih encimov smo merili preko dokazovanja sproščenih reducirajočih sladkorjev, lakazno aktivnost pa preko oksidacije ABTS. Za oceno števila in molekulske mase tarčnih encimov v ekstraktih smo jih ločili tudi na encimogramih. HYUL je med rastjo na mulju izločal vsaj 7 celulolitičnih encimov (20-100 kDa), 6 ksilanolitičnih encimov (5-22 kDa) in 1 encim z lakazno aktivnostjo (10 kDa).

• Language: Slovenian
• Keywords: biotehnologija, lignocelulolitični encimi, glive, papirna industrija, odpadni mulj, encimska aktivnost, Hypsizygus ulmarius, gojenje na trdnem gojišču
• Work type: Master's thesis/paper
• Typology: 2.09 - Master's Thesis
• Organization: BF - Biotechnical Faculty
• Publisher: [N. Jurinčič]
• Year: 2018
• PID: 20.500.12556/RUL-104768
• UDC: 606:61:628.3/.4:604.4:577.15(043.2)
• COBISS.SI-ID: 9085305
• Publication date in RUL: 10.10.2018

Secondary language

• Language: English
• Title: Lignocellulolytic enzyme production on solid waste from pulp and paper industry

Abstract

There is huge amounts of waste sludges produced in pulp and paper industry that imposes significat environmental and economical problem. The purpose of the following work was to show that paper sludge can be used as a substrate for fungi producing lignocellulolytic enzymes. Twenty-nine fungal strains were tested for their ability to grow on paper sludge. Ten different strains (H35, CHP 4, PP3, PLO 4, HYUL, POLY xy, AASC, COCO, GAL 5 in PLO ZABR) that exhibited highest growth-rate on paper sludge were chosen for further analysis. Enzymatic activities in enzyme extracts from selected fungal cultures were measured and three strains were chosen for further experiments. We inoculateted autoclaved (124 °C, 99 min), heat treated (60°C, 180 min) and thermally untreated paper sludge with strains H35, PP3 and HYUL. No growth of any of the strains was observed on paper sludge pretreated by exposure to 60 °C for 180 min. Among the strains able to grow on non-pretreated and autoclaved substrate, best performance was observed with the strain HYUL. Paper sludge overgrown by HYUL was homogenized and crude enzyme extract was analyzed for endoglucanase, xylanase and laccase activities. Number and molecular weights of the lignocellulolytic enzymes in the extracts were estimated on zymograms. During growth on paper sludge, strain HYUL produced at least 7 different cellulolytic enzymes (20-100 kDa), 6 xylanolytic enzymes (5-22 kDa) and 1 enzyme with laccase activity (10 kDa).

• Keywords: biotechnology, lignocellulolytic enzymes, fungi, pulp and paper industry, waste sludge, enzyme activity, Hypsizygus ulmarius, solid-state fermentation