EXPLORATION, PURIFICATION, AND CHARACTERIZATION OF THERMOSTABIL AND ALCOHOL TOLERANT LIPASE FROM COMPOST ISOLATE

Lipase is one of the hydrolase enzymes used as biocatalysts in industries, e.g, food, oil and fat industry, synthesis of flavor compounds, biopolymers and biodiesel. The ability of lipase to catalyze transesterification reactions may be utilized in biodiesel synthesis. The use of lipase as biodiesel...

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Main Author: FAKHOMAH SYIHAB (NIM: 30512013), SYIFA
Format: Theses
Language:Indonesia
Online Access:https://digilib.itb.ac.id/gdl/view/24460
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Institution: Institut Teknologi Bandung
Language: Indonesia
id id-itb.:24460
institution Institut Teknologi Bandung
building Institut Teknologi Bandung Library
continent Asia
country Indonesia
Indonesia
content_provider Institut Teknologi Bandung
collection Digital ITB
language Indonesia
description Lipase is one of the hydrolase enzymes used as biocatalysts in industries, e.g, food, oil and fat industry, synthesis of flavor compounds, biopolymers and biodiesel. The ability of lipase to catalyze transesterification reactions may be utilized in biodiesel synthesis. The use of lipase as biodiesel catalyst requires a thermostable and alcohol tolerant lipase. To meet these criteria, exploration towards thermostable and alcohol tolerant lipase are required. Exploration of lipase from various sources in nature were carried out to obtain lipase with unique characteristics. One of the unique and unexplored environmental sources is compost. The objective of this research is to discover lipase producing bacteria and to isolate thermostable lipase enzymes that are tolerant to alcohol. <br /> <br /> <br /> Compost samples were used as sources to obtain alcohol tolerant lipase producing bacteria. Sampling was carried out at thermophilic phase of composting (72 °C). Identification of bacteria were carried out using gram staining and ribotyping analysis. Lipase production was performed using modified compost media at 55°C. The crude extract of lipase was partially purified using ammonium sulfate and acetone fractionation. Further purification using ion-exchange chromatography and gel filtration were carried out against the protein fraction with highest specific activity. Subsequently, the result from purified acetone fractionation purification were characterized based on their biochemical properties and tolerance against methanol. <br /> <br /> <br /> Selection of lipase-producing microorganisms was resulted five isolates, namely as AL17, AL59, AL64, AL89, and AL96 isolates. Screening of methanol tolerant bacteria showed that AL17 isolate able to grow in medium with highest methanol concentration (5%). Moreover, optimization of growth conditions shows that bacteria able to grow better on compost modified medium (MK) at 55 °C. Optimation towards pH of medium shows that each isolate has different compatibility. Growth curve from five isolates showed that AL89 has the highest growth rate among other isolates. Lipase expression from AL17 only occured at the late of stationary phase, while other isolates express lipase during logarithmic phase and continued until the end of stationary phase. <br /> <br /> <br /> Lipase is one of the hydrolase enzymes used as biocatalysts in industries, e.g, food, oil and fat industry, synthesis of flavor compounds, biopolymers and biodiesel. The ability of lipase to catalyze transesterification reactions may be utilized in biodiesel synthesis. The use of lipase as biodiesel catalyst requires a thermostable and alcohol tolerant lipase. To meet these criteria, exploration towards thermostable and alcohol tolerant lipase are required. Exploration of lipase from various sources in nature were carried out to obtain lipase with unique characteristics. One of the unique and unexplored environmental sources is compost. The objective of this research is to discover lipase producing bacteria and to isolate thermostable lipase enzymes that are tolerant to alcohol. <br /> <br /> <br /> Compost samples were used as sources to obtain alcohol tolerant lipase producing bacteria. Sampling was carried out at thermophilic phase of composting (72 °C). Identification of bacteria were carried out using gram staining and ribotyping analysis. Lipase production was performed using modified compost media at 55°C. The crude extract of lipase was partially purified using ammonium sulfate and acetone fractionation. Further purification using ion-exchange chromatography and gel filtration were carried out against the protein fraction with highest specific activity. Subsequently, the result from purified acetone fractionation purification were characterized based on their biochemical properties and tolerance against methanol. <br /> <br /> <br /> Selection of lipase-producing microorganisms was resulted five isolates, namely as AL17, AL59, AL64, AL89, and AL96 isolates. Screening of methanol tolerant bacteria showed that AL17 isolate able to grow in medium with highest methanol concentration (5%). Moreover, optimization of growth conditions shows that bacteria able to grow better on compost modified medium (MK) at 55 °C. Optimation towards pH of medium shows that each isolate has different compatibility. Growth curve from five isolates showed that AL89 has the highest growth rate among other isolates. Lipase expression from AL17 only occured at the late of stationary phase, while other isolates express lipase during logarithmic phase and continued until the end of stationary phase.
format Theses
author FAKHOMAH SYIHAB (NIM: 30512013), SYIFA
spellingShingle FAKHOMAH SYIHAB (NIM: 30512013), SYIFA
EXPLORATION, PURIFICATION, AND CHARACTERIZATION OF THERMOSTABIL AND ALCOHOL TOLERANT LIPASE FROM COMPOST ISOLATE
author_facet FAKHOMAH SYIHAB (NIM: 30512013), SYIFA
author_sort FAKHOMAH SYIHAB (NIM: 30512013), SYIFA
title EXPLORATION, PURIFICATION, AND CHARACTERIZATION OF THERMOSTABIL AND ALCOHOL TOLERANT LIPASE FROM COMPOST ISOLATE
title_short EXPLORATION, PURIFICATION, AND CHARACTERIZATION OF THERMOSTABIL AND ALCOHOL TOLERANT LIPASE FROM COMPOST ISOLATE
title_full EXPLORATION, PURIFICATION, AND CHARACTERIZATION OF THERMOSTABIL AND ALCOHOL TOLERANT LIPASE FROM COMPOST ISOLATE
title_fullStr EXPLORATION, PURIFICATION, AND CHARACTERIZATION OF THERMOSTABIL AND ALCOHOL TOLERANT LIPASE FROM COMPOST ISOLATE
title_full_unstemmed EXPLORATION, PURIFICATION, AND CHARACTERIZATION OF THERMOSTABIL AND ALCOHOL TOLERANT LIPASE FROM COMPOST ISOLATE
title_sort exploration, purification, and characterization of thermostabil and alcohol tolerant lipase from compost isolate
url https://digilib.itb.ac.id/gdl/view/24460
_version_ 1821844676693458944
spelling id-itb.:244602017-09-27T15:39:50ZEXPLORATION, PURIFICATION, AND CHARACTERIZATION OF THERMOSTABIL AND ALCOHOL TOLERANT LIPASE FROM COMPOST ISOLATE FAKHOMAH SYIHAB (NIM: 30512013), SYIFA Indonesia Theses INSTITUT TEKNOLOGI BANDUNG https://digilib.itb.ac.id/gdl/view/24460 Lipase is one of the hydrolase enzymes used as biocatalysts in industries, e.g, food, oil and fat industry, synthesis of flavor compounds, biopolymers and biodiesel. The ability of lipase to catalyze transesterification reactions may be utilized in biodiesel synthesis. The use of lipase as biodiesel catalyst requires a thermostable and alcohol tolerant lipase. To meet these criteria, exploration towards thermostable and alcohol tolerant lipase are required. Exploration of lipase from various sources in nature were carried out to obtain lipase with unique characteristics. One of the unique and unexplored environmental sources is compost. The objective of this research is to discover lipase producing bacteria and to isolate thermostable lipase enzymes that are tolerant to alcohol. <br /> <br /> <br /> Compost samples were used as sources to obtain alcohol tolerant lipase producing bacteria. Sampling was carried out at thermophilic phase of composting (72 °C). Identification of bacteria were carried out using gram staining and ribotyping analysis. Lipase production was performed using modified compost media at 55°C. The crude extract of lipase was partially purified using ammonium sulfate and acetone fractionation. Further purification using ion-exchange chromatography and gel filtration were carried out against the protein fraction with highest specific activity. Subsequently, the result from purified acetone fractionation purification were characterized based on their biochemical properties and tolerance against methanol. <br /> <br /> <br /> Selection of lipase-producing microorganisms was resulted five isolates, namely as AL17, AL59, AL64, AL89, and AL96 isolates. Screening of methanol tolerant bacteria showed that AL17 isolate able to grow in medium with highest methanol concentration (5%). Moreover, optimization of growth conditions shows that bacteria able to grow better on compost modified medium (MK) at 55 °C. Optimation towards pH of medium shows that each isolate has different compatibility. Growth curve from five isolates showed that AL89 has the highest growth rate among other isolates. Lipase expression from AL17 only occured at the late of stationary phase, while other isolates express lipase during logarithmic phase and continued until the end of stationary phase. <br /> <br /> <br /> Lipase is one of the hydrolase enzymes used as biocatalysts in industries, e.g, food, oil and fat industry, synthesis of flavor compounds, biopolymers and biodiesel. The ability of lipase to catalyze transesterification reactions may be utilized in biodiesel synthesis. The use of lipase as biodiesel catalyst requires a thermostable and alcohol tolerant lipase. To meet these criteria, exploration towards thermostable and alcohol tolerant lipase are required. Exploration of lipase from various sources in nature were carried out to obtain lipase with unique characteristics. One of the unique and unexplored environmental sources is compost. The objective of this research is to discover lipase producing bacteria and to isolate thermostable lipase enzymes that are tolerant to alcohol. <br /> <br /> <br /> Compost samples were used as sources to obtain alcohol tolerant lipase producing bacteria. Sampling was carried out at thermophilic phase of composting (72 °C). Identification of bacteria were carried out using gram staining and ribotyping analysis. Lipase production was performed using modified compost media at 55°C. The crude extract of lipase was partially purified using ammonium sulfate and acetone fractionation. Further purification using ion-exchange chromatography and gel filtration were carried out against the protein fraction with highest specific activity. Subsequently, the result from purified acetone fractionation purification were characterized based on their biochemical properties and tolerance against methanol. <br /> <br /> <br /> Selection of lipase-producing microorganisms was resulted five isolates, namely as AL17, AL59, AL64, AL89, and AL96 isolates. Screening of methanol tolerant bacteria showed that AL17 isolate able to grow in medium with highest methanol concentration (5%). Moreover, optimization of growth conditions shows that bacteria able to grow better on compost modified medium (MK) at 55 °C. Optimation towards pH of medium shows that each isolate has different compatibility. Growth curve from five isolates showed that AL89 has the highest growth rate among other isolates. Lipase expression from AL17 only occured at the late of stationary phase, while other isolates express lipase during logarithmic phase and continued until the end of stationary phase. text