Path: TopS2-ThesesBiotechnology-SITH2017

PRODUKSI ENZIM LAKASE OLEH Marasmius sp. DARI TANDAN KOSONG KELAPA SAWIT MENGGUNAKAN TRICKLE-BED BIOREACTOR

LACCASE PRODUCTION BY Marasmius sp. FROM OIL PALM EMPTY FRUIT BUNCH (OPEFB) USING TRICKLE-BED BIOREACTOR

Master Theses from JBPTITBPP / 2017-09-29 14:20:42
Oleh : FAIZAH KHOIRUNNISAA NIM: 21116025, S2 - Biotechnology-SITH
Dibuat : 2017-09-29, dengan 1 file

Keyword : Aeration rate, laccase, Marasmius sp., oil palm empty fruit bunch, trickle-bed bioreactor

Lakase merupakan enzim yang dapat digunakan dalam berbagai macam proses di

bidang bioteknologi, seperti bioremediasi polutan fenol di lingkungan,

dekolorisasi pewarna pada limbah tekstil, serta pemutihan pulp. Hal tersebut

disebabkan lakase memiliki spesifitas substrat yang luas, bersifat ramah

lingkungan, dan menggunakan oksigen sebagai akseptor elektron terakhir. Enzim

lakase dapat diproduksi oleh jamur pelapuk putih, salah satunya Marasmius sp.

dengan memanfaatkan komponen lignin dari lignoselulosa yang terdapat pada

limbah pertanian, seperti tandan kosong kelapa sawit (TKKS). Pemanfaatan

TKKS tersebut dapat menekan biaya produksi enzim dan menjawab kebutuhan

enzim lakase yang tinggi. Produksi enzim lakase oleh Marasmius sp. dari TKKS

dapat dilakukan menggunakan trickle-bed bioreactor dengan memerhatikan laju

aerasi yang dapat memengaruhi pertumbuhan dan aktivitas metabolisme

Marasmius sp. Penelitian ini bertujuan untuk menentukan laju aerasi optimum

(1,0; 1,5; dan 2,0 vvm) yang mampu memproduksi enzim lakase dengan aktivitas

tertinggi menggunakan trickle-bed bioreactor. Produksi enzim dilakukan

menggunakan TKKS dengan ukuran sebesar 1 cm dan kondisi suhu 32ºC selama

14 hari. Selama produksi enzim, medium Kirk (pH awal 4,5) dialirkan secara

terus-menerus ke dalam bioreaktor dengan laju alir medium 10 mL/menit dan

dilakukan penambahan medium sebanyak 1,5 liter pada hari ke-1 hingga ke-4,

hari ke-7, dan hari ke- 10. Pengambilan sampel dilakukan setiap 12 jam untuk

mengukur aktivitas enzim lakase, lignin peroksidase (LiP), mangan peroksidase

(MnP), dan selulase menggunakan metode spektrofotometri. Selain itu dilakukan

pengukuran pH, kadar lignin, dan gula pereduksi. Hasil penelitian menunjukkan

bahwa aktivitas lakase tertinggi hingga terendah secara berturut-turut dihasilkan

oleh laju aerasi 1,5 vvm (36,66 U/mL pada jam ke-84); 1,0 vvm (80 U/mL pada

jam ke-180); dan 2,0 vvm (19,36 U/mL pada jam ke-84). Selain lakase, aktivitas

enzim selulase, LiP, dan MnP dapat terdeteksi; namun nilainya lebih rendah

dibandingkan aktivitas lakase. Berdasarkan hasil tersebut dapat disimpulkan

bahwa laju aerasi dapat memengaruhi aktivitas lakase dan laju aerasi 1,5 vvm

merupakan laju optimum untuk produksi enzim lakase

Deskripsi Alternatif :

Laccase is an enzyme that can be used in a variety of biotechnological processes,

such as the bioremediation of phenol pollutants, dye decolorization on textile

waste, and pulp bleaching. This is because laccase has a wide substrate specificity,

eco-friendly, and uses oxygen as the final electron acceptor. Laccase enzyme can

be produced by white-rot fungi, i.e. Marasmius sp. by degrading lignin

component of lignocellulose in agricultural waste, such as oil palm empty fruit

bunches (OPEFB). Utilization of OPEFB can reduce enzyme production costs and

answering high enzyme demand. Production of laccase enzyme by Marasmius sp.

from OPEFB can be done using trickle-bed bioreactor with considering the

aeration rate that may affect the growth and metabolic activity of Marasmius sp.

The aim of study is to determine the optimum aeration rate (1,0; 1,5; and 2,0 vvm)

in producing the highest activity of laccase using trickle-bed bioreactor. The

enzyme production was performed using OPEFB with size of 1 cm and a

temperature of 32ºC for 14 days. During enzyme production, the Kirk medium

(initial pH 4,5) was trickled continuously into the bioreactor at a rate of 10

mL/min and 1,5 liters of the medium was added on day 1 to 4, day 7, and day 10.

The effluent is taken every 12 hours for measurement of enzyme activity: laccase,

lignin peroxidase (LiP), manganese peroxidase (MnP), and cellulose using the

spectrophotometric method. In addition; pH, lignin content, and reducing sugars

were also measured. The results showed that the highest to the lowest laccase

activity was produced respectively by the aeration rate of 1.5 vvm (36.66 U/mL at

the 84th hour); 1.0 vvm (23.80 U/mL at the 180th hour); and 2.0 vvm (19.36 U/mL

at the 84th hour). Other than laccase; the activity of cellulase, LiP, and MnP was

detected; but their activity was lower than laccase. Based on these results, it can

be concluded that the aeration rate can affect the laccase activity and the aeration

rate of 1.5 vvm is the optimum rate for laccase production.

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