Institutional Repository | Satya Wacana Christian University: Determination of Rhodopseudomonas Palustris Cell Biomass Based on Protein Spectra of NIR Spectroscopy
Determination of Rhodopseudomonas
palustris cell biomass based on protein
spectra of NIR Spectroscopy
Thesis
Submitted to
Biology Master Program
to Achieve a Master of Science (M. Si)
Mega Novita
NPM : 422009101
Biology Master Program
Postgraduate Program
Satya Wacana Christian University
Salatiga
2011
APPROVAL SHEET
Thesis title
: Determination of
Rhodopseudomonas palustris cell
biomass based on protein spectra
of NIR Spectroscopy
Name of Student : Mega Novita
NPM
: 422009101
Study Program
: Master of Biology
Has been approved by,
Supervisor I
Supervisor II
Dr. Ferdy S Rondonuwu, M.Sc
Drs. Jubhar Mangimbulude, M.Sc
Approval by,
Dean of Magister Biology
Ir. Ferry F. Karwur, M.Sc., Ph.D.
Has graduated on:
APPROVAL STATEMENT FOR THESIS
PUBLICATION OF ACADEMIC
INTEREST
As a Satya Wacana Christian University, I am
Name
NPM
Study Program
Type of Work
:
:
:
:
Mega Novita, S.Si
422009101
Master of Biology
Thesis
For the development of science, agreed to grant nonexclusive royalty free right to Satya Wacana Christian
University on my scientific work entitled:
“Determination of Rhodopseudomonas palustris cell
biomass based on protein spectra of NIR
Spectroscopy”
With this non-exclusive royalty free right, Satya Wacana
Christian University has a right to keeping, changing
the media or reformatting, managing in the data base,
caring ang publishing my thesis as long as putting my
name as author or creator.
Salatiga, April 2011
Mega Novita, S. Si.
Acquaint,
Supervisor I
Supervisor II
Dr. Ferdy S Rondonuwu, M.Sc
Drs. Jubhar Mangimbulude, M.Sc
LETTER OF DECLARATION
I am, Mega Novita, declare that thesis entitled
“Determination of Rhodopseudomonas palustris cell
biomass based on protein spectra of NIR
Spectroscopy” is my original own work. This thesis
contains no plagiarism material, in whole or in part,
which does not attach any references from the original
authors. If I am proven to perform any plagiarism, the
title which has already given to me can be revoked or
canceled
Salatiga, April 2011
Mega Novita, S. Si.
Preface
Praise and thanksgiving to Allah, so that I can finish this
thesis. For all kindess I would like to thank to:
1. Bureau of Planning and International
Cooperation-
Ministry of national Education, Indonesia (BPKLN) for
giving
double
degree
scholarship
through
master
program of Biology at Satya Wacana Christian University
Salatiga and Kwansei Gakuin University Japan.
2. Dr. Ferry Karwur, M. Sc., as a dean of Biology Master
Study Program, for his encouragement and motivation
during in his lecture and any internal scientific events.
3. Dr.
Ferdy
Samuel
Rondonuwu,
M.
Sc.,
for
his
supervision, scientific discussion and advice.
4. Drs. Jubhar Christian Mangimbulude, M. Sc., for his
supervision, scientific discussion and advice.
5. Prof. Dr. Ir. Haryono Semangun as a senior lecturer at
Satya Wacana Chistian University for his supervision,
scientific discussion and advice.
6. Dr. Budhi Prasetyo, M. Biol., for his supervision,
scientific discussion and advice.
7. All staff and students of Biology Master Study Program
and Laboratory, who helped and supported me.
8. My family and my beloved special friend for love and
spirit
to
finish
study
at
University until this end.
i
Satya
Wacana
Chistian
I realize that there are still many shortcomings of this thesis,
however I hope this thesis can be useful.
Mega Novita
ii
List of Contents
1.
Preface…………………………………………………….…
i
2.
List of Contents…………………………………..……….
iii
3.
List of Figures………………………………….………….
iv
4.
Abstract……………………………………………………..
v
5.
Chapter I. Introduction……………….…………………
1
6.
Chapter II. Materials and Methods……………….…..
3
A. Preparation of synthetic medium ……………....
3
B. Pre-culture………………………………................
3
C. Culture………………..…………………………….… 4
D. Measurement by UV-Vis Spectroscopy…….….
4
E. OD standard curve of cell dry weight…………..
4
F. Pigment extraction…………………………........... 5
G. Measurement by NIR Spectroscopy…………….. 6
7.
Chapter III. Results and Discussion…………..……..
7
8.
Chapter VI. Conclusion…………………………..…..…
17
9.
References…………………………..............................
18
iii
List of Figures
Figure 1. Views colony of Rps. Palustris bacterial cells…..
2
Figure 2. Rps. Palustris biomass production………………..
7
Figure 3. Rps. Palustris culture 0th day (left) and 1st day
(right)……………………………………………………
9
Figure 4. Spectrum of photosynthetic pigments
carotenoid and bacteriochlorophyll of Rps.
Palustris culture……………………………...……… 10
Figure 5. Second derivative spectra of Rsp. Palustris
culture using NIR Spectroscopy……………….…. 12
Figure 6. Original spectra of Rps. Palustris culture using
NIR Spectroscopy………………………………….…
14
Figure 7. Second derivative spectra of protein which is
measured by using NIR spectroscopy at 4900
cm-1………………………………………….………….. 14
Figure 8. Relation between protein measured by NIR
spectroscopy and cell biomass………………….... 15
Figure 9. Validation correlation curve of protein
absorption of NIR Spectroscopy and Rps.
Palustris.cell biomass……………………………….. 16
iv
Abstract
Rhodopseudomonas palustris is a very beneficial bacteria to
humans. These bacteria are able to absorb carbon from the
environment, degrade toxic compounds, also produce natural
dyes. Because of its usefulness, many researchers are
interested to do further study. From what they were doing,
biomass becomes an important factor when analyzing the
optimization of the bacteria production. During the years, the
researchers calculate a such productivity using cell biomass
extraction technique which is somewhat time consuming.
From this background, in this study will be correlated both the
Rhodopseudomonas palustris bacteria growth pattern and its
protein spectra that were measured by NIR Spectroscopy.
From this research results, I expect to facilitate the acquisition
of cell biomass without any extraction procedures.
Key words: Rps. palustris, biomass, protein, NIR
v
palustris cell biomass based on protein
spectra of NIR Spectroscopy
Thesis
Submitted to
Biology Master Program
to Achieve a Master of Science (M. Si)
Mega Novita
NPM : 422009101
Biology Master Program
Postgraduate Program
Satya Wacana Christian University
Salatiga
2011
APPROVAL SHEET
Thesis title
: Determination of
Rhodopseudomonas palustris cell
biomass based on protein spectra
of NIR Spectroscopy
Name of Student : Mega Novita
NPM
: 422009101
Study Program
: Master of Biology
Has been approved by,
Supervisor I
Supervisor II
Dr. Ferdy S Rondonuwu, M.Sc
Drs. Jubhar Mangimbulude, M.Sc
Approval by,
Dean of Magister Biology
Ir. Ferry F. Karwur, M.Sc., Ph.D.
Has graduated on:
APPROVAL STATEMENT FOR THESIS
PUBLICATION OF ACADEMIC
INTEREST
As a Satya Wacana Christian University, I am
Name
NPM
Study Program
Type of Work
:
:
:
:
Mega Novita, S.Si
422009101
Master of Biology
Thesis
For the development of science, agreed to grant nonexclusive royalty free right to Satya Wacana Christian
University on my scientific work entitled:
“Determination of Rhodopseudomonas palustris cell
biomass based on protein spectra of NIR
Spectroscopy”
With this non-exclusive royalty free right, Satya Wacana
Christian University has a right to keeping, changing
the media or reformatting, managing in the data base,
caring ang publishing my thesis as long as putting my
name as author or creator.
Salatiga, April 2011
Mega Novita, S. Si.
Acquaint,
Supervisor I
Supervisor II
Dr. Ferdy S Rondonuwu, M.Sc
Drs. Jubhar Mangimbulude, M.Sc
LETTER OF DECLARATION
I am, Mega Novita, declare that thesis entitled
“Determination of Rhodopseudomonas palustris cell
biomass based on protein spectra of NIR
Spectroscopy” is my original own work. This thesis
contains no plagiarism material, in whole or in part,
which does not attach any references from the original
authors. If I am proven to perform any plagiarism, the
title which has already given to me can be revoked or
canceled
Salatiga, April 2011
Mega Novita, S. Si.
Preface
Praise and thanksgiving to Allah, so that I can finish this
thesis. For all kindess I would like to thank to:
1. Bureau of Planning and International
Cooperation-
Ministry of national Education, Indonesia (BPKLN) for
giving
double
degree
scholarship
through
master
program of Biology at Satya Wacana Christian University
Salatiga and Kwansei Gakuin University Japan.
2. Dr. Ferry Karwur, M. Sc., as a dean of Biology Master
Study Program, for his encouragement and motivation
during in his lecture and any internal scientific events.
3. Dr.
Ferdy
Samuel
Rondonuwu,
M.
Sc.,
for
his
supervision, scientific discussion and advice.
4. Drs. Jubhar Christian Mangimbulude, M. Sc., for his
supervision, scientific discussion and advice.
5. Prof. Dr. Ir. Haryono Semangun as a senior lecturer at
Satya Wacana Chistian University for his supervision,
scientific discussion and advice.
6. Dr. Budhi Prasetyo, M. Biol., for his supervision,
scientific discussion and advice.
7. All staff and students of Biology Master Study Program
and Laboratory, who helped and supported me.
8. My family and my beloved special friend for love and
spirit
to
finish
study
at
University until this end.
i
Satya
Wacana
Chistian
I realize that there are still many shortcomings of this thesis,
however I hope this thesis can be useful.
Mega Novita
ii
List of Contents
1.
Preface…………………………………………………….…
i
2.
List of Contents…………………………………..……….
iii
3.
List of Figures………………………………….………….
iv
4.
Abstract……………………………………………………..
v
5.
Chapter I. Introduction……………….…………………
1
6.
Chapter II. Materials and Methods……………….…..
3
A. Preparation of synthetic medium ……………....
3
B. Pre-culture………………………………................
3
C. Culture………………..…………………………….… 4
D. Measurement by UV-Vis Spectroscopy…….….
4
E. OD standard curve of cell dry weight…………..
4
F. Pigment extraction…………………………........... 5
G. Measurement by NIR Spectroscopy…………….. 6
7.
Chapter III. Results and Discussion…………..……..
7
8.
Chapter VI. Conclusion…………………………..…..…
17
9.
References…………………………..............................
18
iii
List of Figures
Figure 1. Views colony of Rps. Palustris bacterial cells…..
2
Figure 2. Rps. Palustris biomass production………………..
7
Figure 3. Rps. Palustris culture 0th day (left) and 1st day
(right)……………………………………………………
9
Figure 4. Spectrum of photosynthetic pigments
carotenoid and bacteriochlorophyll of Rps.
Palustris culture……………………………...……… 10
Figure 5. Second derivative spectra of Rsp. Palustris
culture using NIR Spectroscopy……………….…. 12
Figure 6. Original spectra of Rps. Palustris culture using
NIR Spectroscopy………………………………….…
14
Figure 7. Second derivative spectra of protein which is
measured by using NIR spectroscopy at 4900
cm-1………………………………………….………….. 14
Figure 8. Relation between protein measured by NIR
spectroscopy and cell biomass………………….... 15
Figure 9. Validation correlation curve of protein
absorption of NIR Spectroscopy and Rps.
Palustris.cell biomass……………………………….. 16
iv
Abstract
Rhodopseudomonas palustris is a very beneficial bacteria to
humans. These bacteria are able to absorb carbon from the
environment, degrade toxic compounds, also produce natural
dyes. Because of its usefulness, many researchers are
interested to do further study. From what they were doing,
biomass becomes an important factor when analyzing the
optimization of the bacteria production. During the years, the
researchers calculate a such productivity using cell biomass
extraction technique which is somewhat time consuming.
From this background, in this study will be correlated both the
Rhodopseudomonas palustris bacteria growth pattern and its
protein spectra that were measured by NIR Spectroscopy.
From this research results, I expect to facilitate the acquisition
of cell biomass without any extraction procedures.
Key words: Rps. palustris, biomass, protein, NIR
v