PRODUKSI BIOETANOL DARI AMPAS SAGU (Metroxylon Sp) MELALUI PROSES PRETREATMENT DAN METODE SIMULTANEOUS SACCHARIFICATION FERMENTATION (SSF).
PRODUKSI BIOETANOL DARI AMPAS SAGU (Metroxylon Sp)
MELALUI PROSES PRETREATMENT DAN METODE
SIMULTANEOUS SACCHARIFICATION FERMENTATION (SSF)
Skripsi Sarjana Kimia
Oleh
KHAIRUNNISAH
BP : 0910413084
JURUSAN KIMIA
FAKULTAS MATEMATIKA DAN ILMU PENGETAHUAN ALAM
UNIVERSITAS ANDALAS
PADANG
2013
i
PRODUKSI BIOETANOL DARI AMPAS SAGU (Metroxylon Sp)
MELALUI PROSES PRETREATMENT DAN METODE
SIMULTANEOUS SACCHARIFICATION FERMENTATION (SSF)
Oleh
KHAIRUNNISAH
BP : 0910413084
Skripsi diajukan untuk memperoleh gelar Sarjana Sains
pada Jurusan Kimia Fakultas Matematika dan Ilmu Pengetahuan Alam
Universitas Andalas
JURUSAN KIMIA
FAKULTAS MATEMATIKA DAN ILMU PENGETAHUAN ALAM
UNIVERSITAS ANDALAS
PADANG
2013
ii
iii
ABSTRACT
Bioethanol Production from Sago Waste (Metroxylon sp) with Pretreatment
Process and Simultaneous Saccharification and Fermentation (SSF) Method
By :
Khairunnisah (0910413084)
Marniati Salim, MS*, Elida Mardiah, MS**
*Advisor I, ** Advisor II
Sago waste is one of biomass lignocellulose that potential to produce
bioethanol. Lignocellulose material consist of three major component such as
cellulose, hemicellulose and lignin. The available of lignin become inhibitor for
enzymatic process it must removed by pretreatment. Research for bioethanol
production from sago waste through pretreatment process by using a variation
of alkaline solution; 1% NaOH, 8% NH4OH, 1% NaOH + 4% NH4OH and 1%
NaOH + 8% NH4OH with a ratio solid to liquid 1:10 (w/v) after that, followed by
SSF method involved two kind of fungi like Trichoderma viride strain T1 sk for
saccharification and Saccharomyces cerevisiae for fermentation process. The
optimum conditions for the reduction of samples obtained after pretreatment at
concentration of 1% NaOH + 8% NH4OH at 49,3017% with a long incubation
period of 3 days with a temperature 500C. Result of determination cellulase
enzyme activity from Trichoderma viride strain T1 sk by 0.1% CMC is 0,1144
unit and the highest glucose yield is 921,25 µg/mL from 0,9 grams substrate of
sago waste while the optimum time for the saccharification process obtained in
75 minutes. Ethanol content was analyzed by GC/MS resulted 12.99% (% area)
or equal with 0,38 mL ethanol yield for 168 hours of fermentation .
Keywords : Sago waste, bioethanol, alkaline pretreatment, SSF
viii
DAFTAR ISI
HALAMAN PENGESAHAN ...........................................................................
iii
HALAMAN PERNYATAAN. ..........................................................................
iv
HALAMAN MOTTO DAN PERSEMBAHAN..................................................
v
INTISARI .......................................................................................................
vii
ABSTRACT. ..................................................................................................
viii
UCAPAN TERIMA KASIH. ............................................................................
ix
DAFTAR ISI...................................................................................................
xi
DAFTAR GAMBAR. ......................................................................................
xiv
DAFTAR TABEL. ..........................................................................................
xv
DAFTAR LAMPIRAN. ...................................................................................
xvi
BAB I PENDAHULUAN.................................................................................
1
1.1 Latar Belakang penelitian.......................................................................
1
1.2 Rumusan Masalah .................................................................................
3
1.3 Tujuan Penelitian ...................................................................................
3
1.4 Manfaat Penelitian .................................................................................
4
BAB II. TINJAUAN PUSTAKA
2.1. Tanaman Sagu (Metroxylon sp) .............................................................
5
2.2. Biomassa Lignoselulosa........................................................................
7
2.2.1 Hemiselulosa................................................................................
7
2.2.2 Lignin ............................................................................................
8
2.2.3 Selulosa ........................................................................................ 9
2.3. Pretreatment lignoselulosa ...................................................................... 9
2.4. Produksi Bioetanol dengan SSF. ............................................................. 12
2.5. GC/MS ....................................................................................................
BAB III. METODOLOGI PENELITIAN.............................................................
15
16
3.1 Waktu dan Tempat Penelitian................................................................
..16
3.2 Alat dan Bahan.. ....................................................................................
..16
3.2.1 Alat ............................................................................................
..16
3.2.2 Bahan ........................................................................................
..16
3.3 Pembuatan Reagen................................................................................... 17
xi
3.3.1 Larutan NaOH 1%......................................................................
17
3.3.2 Larutan NH4OH 4%....................................................................
17
3.3.3 Larutan NH4OH 8%....................................................................
17
3.3.4 Nelson .......................................................................................
17
3.3.5 Fosfomolibdat. ...........................................................................
17
3.3.6 Buffer Asam Asetat pH 5............................................................
17
3.3.7 Buffer Sitrat pH 5 .......................................................................
18
3.3.8 Larutan standar glukosa............................................................
18
3.4 Pembuatan Medium
3.4.1 Medium PDA..............................................................................
18
3.4.2 Medium Produksi Enzim .............................................................
18
3.4.3 Medium YPD................... ...........................................................
19
3.5. Prosedur Percobaan
3.5.1 Preparasi Sampel .......................................................................
19
3.5.2 Pretreatment...... .........................................................................
19
3.5.3 Uji Kualitatif Lignin ......................................................................
19
3.5.4 Peremajaan Jamur Trichoderma viride Strain T1 sk...................
19
3.5.5 Produksi Enzim Selulase. ...........................................................
20
3.5.6 Penentuan Aktivitas Enzim dengan Substrat CMC ..................... 20
3.5.7 Sakarifikasi Enzimatik Variasi Jumlah Substrat Ampas Sagu...... 20
3.5.8 Sakarifikasi Enzimatik Variasi Lama Sakarifikasi Ampas Sagu ...
21
3.5.9 Isolasi dan Pemurnian Saccharomyces cerevisiae ..................... 21
3.5.10 Persiapan Inokulum Saccharomyces cerevisiae ....................... 21
3.5.11 Produksi Bioetanol dengan SSF. ..............................................
22
3.5.12 Penentuan Konsentrasi Etanol. ................................................. 22
BAB IV. HASIL DAN PEMBAHASAN
4.1 Pretreatment sampel dengan basa NaOH dan NH4OH .........................
23
4.2 Peremajaan Jamur Trichoderma viride Strain T1 sk ..............................
26
4.3 Sakarifikasi Enzimatik...........................................................................
26
4.3.1 Hasil Aktivitas Enzim dengan Substrat CMC. ...............................
27
4.3.2 Pengaruh Jumlah Substrat Ampas Sagu Terhadap
Kadar Glukosa...............................................................................
xii
27
4.3.3 Pengaruh Lama Sakarifikasi Terhadap Kadar Glukosa................
28
4.4 Isolasi Saccharomyces cerevisiae dari Fermipan ..................................
29
4.5 Bioetanol dengan SSF...........................................................................
29
BAB V. KESIMPULAN DAN SARAN............................................................
33
5.1 Kesimpulan ...........................................................................................
33
5.2 Saran ....................................................................................................
33
DAFTAR PUSTAKA.......................................................................................
34
LAMPIRAN.....................................................................................................
37
BIODATA PENULIS.......................................................................................
51
MELALUI PROSES PRETREATMENT DAN METODE
SIMULTANEOUS SACCHARIFICATION FERMENTATION (SSF)
Skripsi Sarjana Kimia
Oleh
KHAIRUNNISAH
BP : 0910413084
JURUSAN KIMIA
FAKULTAS MATEMATIKA DAN ILMU PENGETAHUAN ALAM
UNIVERSITAS ANDALAS
PADANG
2013
i
PRODUKSI BIOETANOL DARI AMPAS SAGU (Metroxylon Sp)
MELALUI PROSES PRETREATMENT DAN METODE
SIMULTANEOUS SACCHARIFICATION FERMENTATION (SSF)
Oleh
KHAIRUNNISAH
BP : 0910413084
Skripsi diajukan untuk memperoleh gelar Sarjana Sains
pada Jurusan Kimia Fakultas Matematika dan Ilmu Pengetahuan Alam
Universitas Andalas
JURUSAN KIMIA
FAKULTAS MATEMATIKA DAN ILMU PENGETAHUAN ALAM
UNIVERSITAS ANDALAS
PADANG
2013
ii
iii
ABSTRACT
Bioethanol Production from Sago Waste (Metroxylon sp) with Pretreatment
Process and Simultaneous Saccharification and Fermentation (SSF) Method
By :
Khairunnisah (0910413084)
Marniati Salim, MS*, Elida Mardiah, MS**
*Advisor I, ** Advisor II
Sago waste is one of biomass lignocellulose that potential to produce
bioethanol. Lignocellulose material consist of three major component such as
cellulose, hemicellulose and lignin. The available of lignin become inhibitor for
enzymatic process it must removed by pretreatment. Research for bioethanol
production from sago waste through pretreatment process by using a variation
of alkaline solution; 1% NaOH, 8% NH4OH, 1% NaOH + 4% NH4OH and 1%
NaOH + 8% NH4OH with a ratio solid to liquid 1:10 (w/v) after that, followed by
SSF method involved two kind of fungi like Trichoderma viride strain T1 sk for
saccharification and Saccharomyces cerevisiae for fermentation process. The
optimum conditions for the reduction of samples obtained after pretreatment at
concentration of 1% NaOH + 8% NH4OH at 49,3017% with a long incubation
period of 3 days with a temperature 500C. Result of determination cellulase
enzyme activity from Trichoderma viride strain T1 sk by 0.1% CMC is 0,1144
unit and the highest glucose yield is 921,25 µg/mL from 0,9 grams substrate of
sago waste while the optimum time for the saccharification process obtained in
75 minutes. Ethanol content was analyzed by GC/MS resulted 12.99% (% area)
or equal with 0,38 mL ethanol yield for 168 hours of fermentation .
Keywords : Sago waste, bioethanol, alkaline pretreatment, SSF
viii
DAFTAR ISI
HALAMAN PENGESAHAN ...........................................................................
iii
HALAMAN PERNYATAAN. ..........................................................................
iv
HALAMAN MOTTO DAN PERSEMBAHAN..................................................
v
INTISARI .......................................................................................................
vii
ABSTRACT. ..................................................................................................
viii
UCAPAN TERIMA KASIH. ............................................................................
ix
DAFTAR ISI...................................................................................................
xi
DAFTAR GAMBAR. ......................................................................................
xiv
DAFTAR TABEL. ..........................................................................................
xv
DAFTAR LAMPIRAN. ...................................................................................
xvi
BAB I PENDAHULUAN.................................................................................
1
1.1 Latar Belakang penelitian.......................................................................
1
1.2 Rumusan Masalah .................................................................................
3
1.3 Tujuan Penelitian ...................................................................................
3
1.4 Manfaat Penelitian .................................................................................
4
BAB II. TINJAUAN PUSTAKA
2.1. Tanaman Sagu (Metroxylon sp) .............................................................
5
2.2. Biomassa Lignoselulosa........................................................................
7
2.2.1 Hemiselulosa................................................................................
7
2.2.2 Lignin ............................................................................................
8
2.2.3 Selulosa ........................................................................................ 9
2.3. Pretreatment lignoselulosa ...................................................................... 9
2.4. Produksi Bioetanol dengan SSF. ............................................................. 12
2.5. GC/MS ....................................................................................................
BAB III. METODOLOGI PENELITIAN.............................................................
15
16
3.1 Waktu dan Tempat Penelitian................................................................
..16
3.2 Alat dan Bahan.. ....................................................................................
..16
3.2.1 Alat ............................................................................................
..16
3.2.2 Bahan ........................................................................................
..16
3.3 Pembuatan Reagen................................................................................... 17
xi
3.3.1 Larutan NaOH 1%......................................................................
17
3.3.2 Larutan NH4OH 4%....................................................................
17
3.3.3 Larutan NH4OH 8%....................................................................
17
3.3.4 Nelson .......................................................................................
17
3.3.5 Fosfomolibdat. ...........................................................................
17
3.3.6 Buffer Asam Asetat pH 5............................................................
17
3.3.7 Buffer Sitrat pH 5 .......................................................................
18
3.3.8 Larutan standar glukosa............................................................
18
3.4 Pembuatan Medium
3.4.1 Medium PDA..............................................................................
18
3.4.2 Medium Produksi Enzim .............................................................
18
3.4.3 Medium YPD................... ...........................................................
19
3.5. Prosedur Percobaan
3.5.1 Preparasi Sampel .......................................................................
19
3.5.2 Pretreatment...... .........................................................................
19
3.5.3 Uji Kualitatif Lignin ......................................................................
19
3.5.4 Peremajaan Jamur Trichoderma viride Strain T1 sk...................
19
3.5.5 Produksi Enzim Selulase. ...........................................................
20
3.5.6 Penentuan Aktivitas Enzim dengan Substrat CMC ..................... 20
3.5.7 Sakarifikasi Enzimatik Variasi Jumlah Substrat Ampas Sagu...... 20
3.5.8 Sakarifikasi Enzimatik Variasi Lama Sakarifikasi Ampas Sagu ...
21
3.5.9 Isolasi dan Pemurnian Saccharomyces cerevisiae ..................... 21
3.5.10 Persiapan Inokulum Saccharomyces cerevisiae ....................... 21
3.5.11 Produksi Bioetanol dengan SSF. ..............................................
22
3.5.12 Penentuan Konsentrasi Etanol. ................................................. 22
BAB IV. HASIL DAN PEMBAHASAN
4.1 Pretreatment sampel dengan basa NaOH dan NH4OH .........................
23
4.2 Peremajaan Jamur Trichoderma viride Strain T1 sk ..............................
26
4.3 Sakarifikasi Enzimatik...........................................................................
26
4.3.1 Hasil Aktivitas Enzim dengan Substrat CMC. ...............................
27
4.3.2 Pengaruh Jumlah Substrat Ampas Sagu Terhadap
Kadar Glukosa...............................................................................
xii
27
4.3.3 Pengaruh Lama Sakarifikasi Terhadap Kadar Glukosa................
28
4.4 Isolasi Saccharomyces cerevisiae dari Fermipan ..................................
29
4.5 Bioetanol dengan SSF...........................................................................
29
BAB V. KESIMPULAN DAN SARAN............................................................
33
5.1 Kesimpulan ...........................................................................................
33
5.2 Saran ....................................................................................................
33
DAFTAR PUSTAKA.......................................................................................
34
LAMPIRAN.....................................................................................................
37
BIODATA PENULIS.......................................................................................
51