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Sakti Imam Muchlissin
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ISOLASI BAKTERI PENGHASIL ENZIM PROTEASE STAPHYLOCOCCUS HOMINIS PADA ONCOM MERAH PASCA FERMENTASI 120 JAM Aulia Harun; Sakti Imam Muchlissin; Ana Hidayati Mukaromah; Sri Darmawati; Stalis Norma Ethica
PROSIDING SEMINAR NASIONAL & INTERNASIONAL 2018: SEMINAR NASIONAL PENDIDIKAN SAINS DAN TEKNOLOGI
Publisher : Universitas Muhammadiyah Semarang

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Abstract

Enzymes are complex protein moleculer produced by living cells playing role as catalysts in various chemical processes in the body. Among enzymes playing an important role in human life is protease. The purpose of this study was to determine the presence of protease – producing bacteria found on 120-h post - fermented oncom and to identify the bacteria based on its 16S   rRNA gene analysis. Bacterial isolation and purification was carried out using Nutrient Agar media with spread technique. Of the six bacterial isolates isolated from the oncom sample after 120 hours of fermentation, there was one isolate that had protease activity, namely IROD 5. The protease enzyme income test was carried out using Skim Milk Agar media. Molecular identification process was carried out through sequential analysis of 16S rRNA using PCR method using primers forward F: 5'-AGAGTTGATCCTGGCTCAG-3 'and reverse R: 5'- GGTTACCTTGTTAC. GACTT-3 primers' followed by sequencing process. The protease enzyme production test to bacterial isolate was conducted using Skim Milk Agar. Molecular identification was performed through analysis of 16S rRNA gene sequence using PCR method followed by sequencing process. A single bacterial isolate having proteolytic activity was obtained based on observation of the clear zone of protease surrounding the bacterial colony with a diameter of 72 mm. The 16S rRNA gene sequence of the obtained proteolytic bacterial strain IROD5 has been obtained and analysis on the gene sequence resulted 99% similarity levels with sequence of similar gene s of Staphylococcus hominis. As conclusion, the obtained bacterial isolate in this studyis apotential protease  enzyme  producer  and  molecularly  identified  as  Staphylococcus hominis strains IROD5. Keyword : Protease Enzyme, Gen 16S rRNA, Red Oncom
ISOLASI BAKTERI PENGHASIL ENZIM PROTEASE BACILLUS THURINGIENSIS IRODI PADA ONCOM MERAH PASCA FERMENTASI 24 JAM Radna Safitri; Sakti Imam Muchlissin; Ana Hidayati Mukaromah; Sri Darmawati; Stalis Norma Ethica
PROSIDING SEMINAR NASIONAL & INTERNASIONAL 2018: SEMINAR NASIONAL PENDIDIKAN SAINS DAN TEKNOLOGI
Publisher : Universitas Muhammadiyah Semarang

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Abstract

The need for protease enzymes in Indonesia and the world continues to increase, requiring  new  protease  sources.  Bacteria  are  beneficial  sources  of  protease because they are easy to obtain and rapidly multiply. Bacterial identification could be done molecularly through analysis of the 16S rRNA gene. This study aimed to obtain an isolate of protease-producing bacterium from 24-h post-fermented red oncom and to identify the obtained bacterial strain molecularly by 16S rRNA gene sequence. The protease production test on bacteria found in red oncom sample was done  using  a  selective  medium,  Skim  Milk  Agar  (SMA).  DNA  genomes  of proteolytic bacterial cells were extracted by Promega KIT. The amplifying process of 16S rRNA gene using the Polymerase Chain Reaction (PCR) method. The amplified  DNA  were  analyzed  using  the  BLAST  program.  The  results  of  the research found 8 isolate of bacterias. The most unique isolate was IROD1.3. It has significant  proteolytic  activity  based  on  the  ability  to  produce  clear  zone  of protease on SMA medium with a diameter of 85,00 mm. Isolate IROD1.3 was identified  molecularly  as  Bacillus  thuringiensis  with  similarity  of  96% to  the sequence of 16S rRNA gene Bacillus thuringiensis strain TERI SID4 (Genbank access code: KX822158.1). Keywords: Protease, 16S ribosomal RNA gene, red oncom, proteolytic bacterium
ISOLASI BAKTERI PENGHASIL ENZIM PROTEASE BACILLUS MEGATERIUM IROD3 DARI ONCOM MERAH PASCA FERMENTASI 72 JAM Dhea Ayu Lestari; Sakti Imam Muchlissin; Ana Hidayanti Mukaromah; Sri Darmawati; Stalis Norma Ethica
PROSIDING SEMINAR NASIONAL & INTERNASIONAL 2018: SEMINAR NASIONAL PENDIDIKAN SAINS DAN TEKNOLOGI
Publisher : Universitas Muhammadiyah Semarang

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Abstract

Protease  enzyme  has  function  to  hydrolyze  peptide  bonds  in  proteins  into simpler molecules to digest by the body which important to food industry. One of effort to increase the production of protease enzymes is looking for new sources of protease particularly from bacterial groups. The purpose of this study was to obtain an isolate of protease-producing bacteria found on post- fermentation oncom 72 hours, and to identify the protease-producing bacteria based on the analysis of 16S rRNA gene. Isolation and purification process of bacterial colony was carried out on Nutrient Agar medium with spread technique, production test of protease enzyme was performed using Selective Skim Milk Agar. The process of Molecular identification process was carried out  through  analysis  of  16S  rRNA  gene  fragment  sequences  which  were amplified using Polymerase Chain Reaction (PCR) method, and continued by sequencing. The result of bacteria isolation was found one isolate which has proteolytic activity in Skim Milk Agar medium which has clear zone diameter of 78.00 mm. A similarity analysis based on the 16S rRNA gene sequence showed that IROD3 (Indonesian Red Oncom Day-3) has 99% similarity level with the 16S rRNA gene fragment of Bacillus megaterium strain CS17 (access code Genbank: MG430224.1). Keywords:  Molecular  identification,  proteolytic  bacteria,  16S  rRNA  gene, Bacillus megaterium
ISOLASI DAN IDENTIFIKASI MOLEKULER BAKTERI PENGHASIL ENZIM PROTEASE PSEUDOMONAS STUTZERI ISTD4 DARI TEMPE GEMBUS PASCA FERMENTASI 1 HARI Wa Ode Inayatul; Sakti Imam Muchlissin; Ana Hidayati Mukaromah; Sri Darmawati; Stalis Norma Ethica
PROSIDING SEMINAR NASIONAL & INTERNASIONAL 2018: SEMINAR NASIONAL PENDIDIKAN SAINS DAN TEKNOLOGI
Publisher : Universitas Muhammadiyah Semarang

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Abstract

Proteaseis  a group  of enzymes that  play  an important  role in  biochemical reactions, whichc a use protein break down. Protease is among main enzymes used in industry, which commercial value reach 60% of total enzymes world wide. This study a imed toisolat protease-producing bacterium found on tempe gembus in after 1-day post-fermentation and to identify the bacterial isolat obtained based on the analysis of its 16S rRNA gene. Isolati on and purification process wasd one using Nutrient Agar media with spread technique. The protease production test was carried out on skim milk agar medium. The molecular identification process was performed by analyzing sequence of 16S rRNA gene fragment of bacteria amplified using both forward primer F (F:5'- AGAGTTGATCCTGGCTCAG-3'), and reverse primer R (R:5'- GGTTACCTTGTTACGACTT-3') by Polymerase Chain Reaction (PCR) method. The amplified DNA from PCR was then sequenced. From the isolation process a bacterial strain that has a proteolytic activity based on observation of clear zone area with a diameter of 85 mm was obtained. From sequence alignment result using BLAST (Basic Local Alignment Search Tool) the fragment of 16S rRNA gene of strain ISTD1.4 obtained has similarity level of 98% with fragment of 16S ribosomal RNA gene of bacterium Pseudomonas stutzeri strain E141. In conclusion, strain ISTD1.4 is a potential protease-producing bacteria and is identified as Pseudomonas stutzeri ISTD4. Keywords: Bacterial isolation, molecular identification, proteolytic bacteria, 16S rRNA gene
ISOLASI BAKTERI PENGHASIL ENZIM PROTEASE BACILLUS AMYLOLIQUEFACIENS IROD2 PADA ONCOM MERAH PASCA FERMENTASI 48 JAM Dwi Pamaya; Sakti Imam Muchlissin; Endang Tri Wahyuni Maharani; Sri Darmawati; Stalis Norma Ethica
PROSIDING SEMINAR NASIONAL & INTERNASIONAL 2018: SEMINAR NASIONAL PENDIDIKAN SAINS DAN TEKNOLOGI
Publisher : Universitas Muhammadiyah Semarang

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Abstract

Proteolytic  bacteria  are  the  bacteria  capable  of  producing  extracellular protease enzymes, namely protein-breaking enzymes that are widely used in many industrial fields. This study aimed to isolate a proteolytic bacterium found on 48-h post-fermented oncom and molecular identification method. The initial isolation and purification process of the colony was carried out using Nutrient Agar medium. Selection of protease enzyme obtained by bacterial isolate was done on Milk Skim Agar medium. Identification process of the isolate was done through amplification of 16S rRNA gene using PCR, sequencing and analysis of gene sequences using BLAST program. From the isolation process  a bacterial isolate that has proteolytic by the ability to produce a clear zone of 82.00 on plate. The result of the 16S rRNA gene sequence analysis showed that the proteolytic bacterial isolate obtained in this study had a 98% homology level with 16S ribosomal RNA isolate of Bacillus amyloliquefaciens strain A1142 (Genbank access code: KTT722836.1). Based on the results of the molecular identification, the isolate was identified as Bacillus amyloliquefaciens strain IROD2  (IROD2 = Indonesia Red Oncom Day2). As conclusion, from 48-h post fermented red oncom, a protease producing bacterial strain molecularly identified as Bacillus amyloliquefaciens strain IROD2. Keywords: Moleculary was identified, Proteolitic bacteria, 16S rRNA gene
SAMPLING MIKROBIOLOGI LIMBAH BIOMEDIS RUMAH SAKIT DI KOTA SEMARANG JAWA TENGAH Stalis Norma Ethica; Sakti Imam Muchlissin; Ragil Saptaningtyas; Agus Sabdono
PROSIDING SEMINAR NASIONAL & INTERNASIONAL 2017: Prosiding Seminar Nasional Publikasi Hasil-Hasil Penelitian dan Pengabdian Masyarakat
Publisher : Universitas Muhammadiyah Semarang

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Abstract

The present study aimed to propose a technical strategy of sampling for obtaining microbiological samples from liquid biomedical wastes generated by hospitals in Semarang City, Central Java Province, Indonesia.  The samples will be used to evaluate bioremediation ability of the hydrolytic bacteria isolated from the waste. It was hoped that the proposed strategy could help to guide researchers about appropriate sampling practices on hospital biomedical wastes from IPAL of Indonesian hospitals. As mandated by the Indonesian law, the handling of hospital liquid waste in Indonesia should follow Permenkes No.1204/2004, while the liquid waste standard should meet the Kepmen LH No.58/1995, reaffirmed by Perda Provinsi Jawa Tengah No.10/2004 about liquid waste standard issued by the Governor of Central Java. Based on formal regulations and safety aspects, it is proposed that the strategy for a proper microbiological sampling practice in terms of hospital liquid biomedical waste should include: (1) Permit letter from hospital where sampling location is; (2) Relevant sampling method based on research purpose (3) Standard, anti-leaked, properly labelled sampling containers(4) Proper sampling equipment based on sampling purpose (5) Adequate sampling transport equipment; (6) The use of standard personal protection equipment (PPE); and (7) Vaccine for the sampling workers (hepatitis B vaccination ismandatory).  Keywords: microbiological sampling, hospital biomedical waste, microbial bioremediation, sampling strategy,Indonesian sampling regulation
Co-Authors Agus Sabdono Ana Hidayanti Mukaromah Ana Hidayati Mukaromah Aulia Harun Dhea Ayu Lestari Dwi Pamaya Endang Tri Wahyuni Maharani Radna Safitri Ragil Saptaningtyas Sri Darmawati Stalis Norma Ethica Wa Ode Inayatul