Articles

ISOLASI DAN UJI AKTIVITAS KITINASE ISOLAT BAKTERI DARI KAWASAN GEOTERMAL DIENG Nafisah, Hidayatun; Pujiyanto, Sri; Raharjo, Budi
Bioma : Berkala Ilmiah Biologi Vol. 19, No. 1, Tahun 2017
Publisher : Departemen Biologi, Fakultas Sains dan Matematika, Universitas Diponegoro

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (201.301 KB) | DOI: 10.14710/bioma.19.1.22-29

Abstract

Chitinase (EC.3.2.2.14) is an enzyme which can degradatechitin became N-acetilglucosamin. Chitinase has many benefits made the demand of it increases. High demands spur its availability in large quantities, cheap, fast production, resistant to any physical factor and chemical environment. Rapid and resistant enzyme production to environment factor can be obtained using chitinolitic bacteria of Geothermal Dieng. The utilization of chitin as bacterial growth substrate from waste of shell crab can be done considering high prices of commercial chitin on the market. The purpose of the research is to get the isolate of termoleranchitinolitic of watery mud in Geothermal Dieng and to know the character of the chosen isolate producing highest chinitase activity type of chitin source treatment and pH of media production. The research is done by growing the chitinolitic in the room temperature for 14 days. The experimental design used in this study is a complete randomized design of factorial pattern (two factors). The first factor is the type of chitin source that includes commercial chitin and chitin crab kits. The second factor is the pH of liquid chitin media for the production of enzymes, ie pH 6, 7 and 8.Chitinase activity is tested by measuring the result of sugar reduction. Obtained data is analyzed with Analysis of Variance (ANOVA). Result of isolation and selection is obtained one potential isolate, KSR 121. The isolate produce 1,4 cm of chitinolitic index after 96 hour incubation. Result of statistical test show both citin source type, pH of media production treatment and interaction were not significantly different (P?0,05). KSR 121 isolate experience the highest growth of crab chitin treatment pH 8 (K2P3) with 6 hour incubation, whereas highest kinitase activity happen on crab chitin treatment pH 7 (K2P2) with 24 incubation, in amount of 0,125 (U/mL). Key words: N-acetil glucosamin, chtinase activity, chitinase, chitin, chitinolitic bacteria, isolation
PRODUKSI PROTEASE ALKALIS TERMOSTABIL DARI ASPERGILLUS FLAVUS DUCC- K225 DENGAN AMMONIUM SULFAT SEBAGAI SUMBER NITROGEN Putra, Mohammad Affan Dwica; Rukmi, MG Isworo; Pujiyanto, Sri; Mulyani, Nies S
Bioma : Berkala Ilmiah Biologi Vol. 21, No 1, Tahun 2019
Publisher : Departemen Biologi, Fakultas Sains dan Matematika, Universitas Diponegoro

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (108.467 KB) | DOI: 10.14710/bioma.21.1.60-64

Abstract

Protease  is a protein hydrolytic enzyme which  can be generated by a variety of microorganisms, including  mold. Aspergillus flavus K225, DUCC is indigenous mold isolated from lime soil of Madura which is  have potential as a alkaline protease  producer. This research aims was to know the effect  of ammonium sulfate as nitrogen source for the production of protease enzymes by Aspergillus flavus DUCC-K225. The production of alkaline  protease were conducted in  submerged culture medium with agitation. Fermentation medium used was modification Czapeks Dox Broth containing 2% casein. Incubation is carried out for 7 days.  The results showed that ammonium sulfate is a good source of nitrogen for growth and production of aklaine protease enzyme, which is demonstrated by  higher dry weight, the protease activity, the protein content and the specific activities, comparing those  on standard medium using sodium nitrate as N source. 
UJI AKTIVITAS KITIN DEASETILASE ISOLAT BAKTERI DARI KAWASAN GEOTERMAL DIENG Akhsani, Ghaida Afra; Suprihadi, Agung; pujiyanto, Sri
Jurnal Akademika Biologi Vol. 6 No. 3 Juli 2017
Publisher : Departemen Biologi, Fakultas Sains dan Matematika Undip

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Abstract

Chitinolytic bacteria is a bacterium, which is able to degrade chitin. This ability is obtained from the resulted chitinolytic enzyme. Chitin deacetylase (EC 3.5.1.41) is one of the chitinolytic enzymes, which be able to convert chitin into its derivatives. For this reason, chitin deacetylase has a chance to be an environmentally enzymatic converter of chitin. In addition, chitin derivatives have a wider potential in many fields. The objectives of this study were to obtain bacterial isolates from the mud of Sikidang Crater in Dieng geothermal field that producing chitin deacetylase enzyme, and to determine its activity characteristic of (optimum time production, optimum pH, and effect of 1 mM divalent metal ions) from the resulted chitin deacetylase enzyme. This research used completely randomized design. The data were analyzed using One Way ANOVA and Tukey HSD test. The results showed that KSR HA 24 isolates were able to produce chitin deasetylase with optimum enzyme activity of 0.668 U / ml at 18 hours production time. Optimum activity of chitin deacetylase occurred at pH 5 of 0.75 U / ml. Chitin deacetylase activity with 1 mM addition of divalent metal ions produce activator metal ions, including Mg2+, which increased the activity up to 154.43%, Fe2+ the activity up to 144.63%, and Cu2+ the activity up to 110.41%. Inhibitor metal ions, including Zn2+, which decreased the activity to 93.77%, and Mn2+ the activity to 86.46%.Keywords: Chitinolytic, Chitin Deacetylase, Enzyme Activity, pH, Divalent Metal Ions
PRODUKSI ENZIM PROTEASE DARI A.NIGER PAM18A DENGAN VARIASI PH DAN WAKTU INKUBASI Ramadhani, Putri; Rukmi, MG. Isworo; pujiyanto, Sri
Jurnal Akademika Biologi Vol. 4 No. 2 April 2015
Publisher : Departemen Biologi, Fakultas Sains dan Matematika Undip

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Abstract

Advances in the field of fermentation technology, genetic engineering, and technology applications of enzymes lead to the use of enzymes in the industry is increasing. Enzymes used in the industry can be derived from microorganisms such as bacteria, molds, and yeasts. Protease is one of the important enzyme that has been used widely in various fields of industrial application and 65% of the sales of enzymes in the world. Alkaline protease is one of hydrolytic enzymes that can hydrolysis proteins which working at the pH range of 7-12. The aim of this study was  to investigate the influence of pH and incubation time on the protease production of A. niger PAM 18A. The experimental were done using Factor Block Random Design with 2 i.e pH of7, 8, 9, and incubation time of 5, 6, 7 days. The results showed that highest protease fromA. niger PAM18A were found in the treatment of pH 8 and of 5 days incubating with the protease activity and specific activity were 0,930 U/mL and 3,632U/mg respectively.Keywords: Production Enzyme, Protease, A. niger PAM18A, pH, incubation time.
UJI AKTIVITAS ANTIBAKTERI EKSTRAK BIJI KELOR (MORINGA OLEIFERA L.) TERHADAP PERTUMBUHAN BAKTERI STAPHYLOCOCCUS AUREUS DAN BAKTERI ESCHERICHIA COLI Wigunarti, Anggia Hesti; Pujiyanto, Sri; Suprihadi, Agung
Berkala Bioteknologi Volume 2 Nomor 2 November 2019
Publisher : Berkala Bioteknologi

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Abstract

Moringa oleifera L. or moringa is one of the plants that can be used as a traditional herbal medicine because it has antimicrobial, anti-tumor, anti-pyretic, anti-oxidant and anti-inflammatory properties. The purpose of this study was to determine the antimicrobial activity of Moringa seed extract against Staphylococcus aureus and Escherichia coli. Extraction of active compounds from moringa seeds was carried out by maceration method. The maceration process with two solvents, polar solvent using 96% ethanol, and second with non-polar solvents using n-hexane. The macerate results are then evaporated in 60oC temperature over 1 hour . The test is done by diffusion to use a 6 mm paperdisk with two test of bacteria Staphylococcus aureus and Escherichia coli. The concentration of extract solution to be tested are 75%, 50% and 25%. The positive control was using 0,1 mg / mL chloramphenicol and negative control using DMSO. Experimental design using a completely randomized design and analyzed using the Analysis of Variance (ANOVA) and if the data obtained showed significantly different results at the 5% test level then continued with the Duncan test to determine differences between the treatments. From the results of testing the antimicrobial activity of Moringa seed extract in 96% polar ethanol solvent showed the presence of antibacterial activity against both of the test bacteria Staphylococcus aureus and Escherichia coli. The highest antibacterial activity occurs at a concentration of 75%. The diameter of the inhibitory zone of Staphylococcus aureus and Escherichia coli was 14,75 and 3,50. Non-polar n-hexane solvents showed no antibacterial activity in the two test bacteria Staphylococcus aureus and Escherichia coli. Key words:Moringa oleifera L., extracts, antibacterial, S.aureus, E.coli
SKRINING AKTIVITAS ANTIBAKTERI DAN IDENTIFIKASI MOLEKULER BERDASARKAN GEN 16S RRNA ISOLAT AKTINOMISET ASAL PULAU ENGGANO DAN BALI Nadina, Rahmah Qisti; Pujiyanto, Sri; Wijanarka, W; Fahrurrozi, Fahrurrozi
Berkala Bioteknologi Volume 2 Nomor 2 November 2019
Publisher : Berkala Bioteknologi

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Actinomycetes are a Gram positive bacteria who plays an important role in pharmaceutical industry because of its ability to produce an antibacterial compound. This study aims to select actynomycetes isolates that has antibacterial activity and to identify the isolates based on 16S rRNA gene squence. The  actinomycetes isolates were inoculated using ISP-2 medium and cultured in SYP (starch, yeast, peptone) media for 13 days. The cultures were extracted using etyl acetate solvent and antibacterial activity was tested against Bacillus subtilis and Escherichia coliusing agar diffusion method.Positive isolates were identified based on 16S rRNA gene sequence. The 16S rRNA gene from positive isolates was sequenced and analyzed with computerized help and was deposited at NCBI. The antibacterial activity test revealed 5 from 22 isolates had an antebacterial activity shown by the clear zone around the whatman filter paper on both tested bacteria. The identification revealed these five isolates were identified asStreptomyces mutabilis, Streptomyces sp, and Streptomyces griseorubens. This study shows these isolates from Enggano and Bali Island are potential as antibacteri producer.  Keywords : actynomycetes, antibacterial activity, 16S rRNA
ANALISIS FUSAN HASIL FUSI PROTOPLAS INTRASPESIES PICHIA MANSHURICA DUCC-015 (ANALYSIS OF FUSANT FROM PROTOPLAST FUSION INTRASPECIES OF PICHIA MANSHURICA DUCC-015) Roslenawati, Roslenawati; Kusumaningrum, Hermin Pancasakti; Pujiyanto, Sri
JURNAL SAINS DAN MATEMATIKA Volume 22 Issue 1 Year 2014
Publisher : JURNAL SAINS DAN MATEMATIKA

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Abstract

The intraspecies protoplast fusion of P. manshurica DUCC-015 was conducted in searching the fusant with greater inulinase production. Inulinase on Dahlia variabilis Willd tuber from Baturraden-Purwokerto showed inulinase activity 0,683 IU/mL. Inulinase is an enzyme that catalyzes the hydrolysis reaction of inulin polysaccharides into fructose and or fructooligosacarides. The aims of this research was revealing fusant from intraspecies protoplast fusion process of P. manshurica DUCC-015 followed by analyzing of their inulinase productivity and activity. The research metodology protoplast fusion process consist of lysis of cell wall, protoplast fusion and regeneration of intraspecies fusant P. manshurica DUCC-015. Analysis of fusant were done by Schiff Basic Fuchsin staining of fusant nuclei, also measuring the inulinase activity and inulinase production comparing with their parent. The inulinase activity will be analyzed by T-Test Independent Two Sample on 95% Confidence interval of the difference use Statistical Product and Service Solution programme (SPSS). The result of experiment gaining fusant with regeneration capability, ploidi diversity  of fusant, inulinase activity about 0,965 IU/mL while their parent 0,622 IU/mL and inulinase production 0,736  IU/mL comparing with 0,731 IU/mL during 42 hours incubations. The fusant indicated the increase of inulinase activity and production  compared with their parent.   Keywords: Pichia manshurica DUCC-015, fusant, inulinase    Fusi protoplas intraspesies Pichia manshurica DUCC-015 telah dilakukan untuk mencari fusan dengan produksi inulinase yang lebih tinggi. Inulinase pada umbi tanaman Dahlia variabilis Willd dari Baturraden-Purwokerto memperlihatkan aktivitas sebesar 0,683 IU/mL. Inulinase merupakan enzim yang mengkatalisis reaksi hidrolisis polisakarida inulin menjadi fruktosa dan atau fruktooligosakarida. Penelitian ini bertujuan untuk mendapatkan fusan hasil fusi protoplas intraspesies P. manshurica DUCC-015 yang memiliki aktivitas inulinase lebih tinggi. Rancangan percobaan fusi protoplas terdiri dari isolasi protoplas, fusi protoplas dan regenerasi fusan. Analisis fusan menggunakan pewarnaan  Fuchsin pada inti sel, mengukur aktivitas dan produksi inulinase fusan. Aktivitas inulinase dianalisis dengan Uji T Test Dua Sampel Independen pada taraf kepercayaan 95% menggunakan program Statistical Product and Service Solution (SPSS). Hasil penelitian menunjukkan fusi protoplas intraspesies P. manshurica     DUCC-015 menghasilkan aktivitas inulinase mencapai 0,965 IU/mL dibandingkan induk sebesar 0,622 IU/mL dan produksi inulinase 0,736  IU/mL pada inkubasi selama 42 jam. Fusan  mengindikasikan kenaikan aktivitas dan roduksi inulinasi dibandingkan induk. Kata kunci : Pichia manshurica DUCC-015, fusan, inulinase.
PRODUKSI DAN EKSTRAKSI INHIBITOR ALFA GLUKOSIDASE DARI ISOLAT AKTINOMISET JP-3 Pujiyanto, Sri; Ferniah, Rejeki Siti; S, Sunarno
Bioma : Berkala Ilmiah Biologi Vol. 17, No.2, Tahun 2015
Publisher : Departemen Biologi, Fakultas Sains dan Matematika, Universitas Diponegoro

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (102.608 KB) | DOI: 10.14710/bioma.17.2.123-129

Abstract

Alpha-glucosidase inhibitors are compounds that can prevent the digestion of complex carbohydrates into glucose, so potentially used as a diabetes drug. This study aims to examine the production and extraction of alpha-glucosidase inhibitor compound from Isolate Aktinomiset JP-3 from the sea. The supernatant obtained from the culture of the JP-3 isolate was extracted using various solvents to obtain the active compound. The solvents used were chloroform, methanol, and ethyl acetate. An assay of inhibitor activity of the ?-glucosidase using p-nitrophenyl ?-D-glucopyranoside substrate. The activity of the enzyme is measured based on the absorbance of p-nitrophenol produced from the breaking reaction of the substrate. The results showed that extraction of alpha-glucosidase inhibitor compound with ethyl acetate yielded extract with highest inhibitor activity. Keywords: alpha-glucosidase inhibitors, actinomycetes, diabetes, extraction, fractionation
AKTIVITAS BAKTERI KITINOLITIK AKUATIK ISOLAT LOKAL TERHADAP PERKEMBANGAN DAN MORTALITAS LARVA NYAMUK AEDES AEGYPTI L Pujiyanto, Sri; Ferniah, RS; Rahardian, R.
JURNAL SAINS DAN MATEMATIKA Volume 19 Issue 2 Year 2011
Publisher : JURNAL SAINS DAN MATEMATIKA

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Abstract

Pengendalian populasi nyamuk Aedes aegypti sangat penting dalam rangka pencegahan terjadinya wabah penyakit demam berdarah. Beberapa bakteri memiliki aktivitas kitinolitik sehingga berpotensi digunakan sebagai agen biokontrol terhadap nyamuk Aedes aegypti, karena komponen eksoskeleton larva nyamuk tersebut tersusun dari bahan kitin.Tujuan dari penelitian ini adalah untuk mendapatkan isolat bakteri kitinolitik akuatik lokal yang dapat digunakan sebagai biokontrol larva nyamuk Aedes aegypti.  Bakteri kitinolitik diisolasi menggunakan media selektif agar kitin. Sumber isolat diperoleh  dari berbagai sampel air yang diperoleh dari daerah Klaten, Boyolali dan Jepara. Isolat yang diperoleh diseleksi untuk mendapatkan isolat dengan aktivitas tinggi. Uji aktivitas bakteri terhadap larva nyamuk menggunakan media air mineral. Hasil penelitian diperoleh satu isolat dengan kode B6 yang mampu menyebabkan kematian larva nyamuk sebesar 97% dalam waktu 108 jam. Isolat ini berpotensi digunakan sebagai agen  bioinsektisida untuk pengendalian larva nyamuk A. aegypti. Keywords: Bakteri kitinolitik, demam berdarah, Aedes aegypti, biokontrol
UJI AKTIVITAS ANTIBAKTERI EKSTRAK TUMBUHAN EUPHORBIA HIRTA L. TERHADAP RALSTONIA SOLANACEARUM, ESCHERICHIA COLI, DAN STAPHYLOCOCCUS AUREUS SECARA IN VITRO Angelika, Genoveva Preta; Suprihadi, Agung; pujiyanto, Sri
Jurnal Akademika Biologi Vol. 3 No. 2 April 2014
Publisher : Departemen Biologi, Fakultas Sains dan Matematika Undip

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Abstract

Biocontrol using Patikan kebo (Euphorbia hirta L.) plant is an alternative solution to control pathogenic bacteria. Such wild plant is known to contain active compounds with antibacterial activity such as tannins, alkaloids, flavonoids, saponins, and phenols. This study aims to determine the antibacterial activity of the methanol extract of E. hirta against R. solanacearum, E. coli and S. aureus. The extraction method of E. hirta was maceration with methanol solvent, while antibacterial activity test using the agar diffusion method (Kirby-Bauer) with test bacteria was R. solanacearum, E. coli and S. aureus. E. hirta extract tested was pure extract (100%). Observed response was diameter of inhibitory zone formed around the paper discs that had been smeared with E. hirta extract on the media. Analysis of the data using a Completely Randomized Design (CRD) 1 factor (test bacteria) with three times repetition, followed by a further test of Duncan with 95% confidence level. The results indicated that E. hirta produced extraction yield of 6,45%. Antibacterial activity of E. hirta extract against R. solanacearum, E. coli and S. aureus was indicated by Inhibitory Zone Diameter (HZD), respectively for 21,8 mm, 18,26 mm and 17,06 mm. The results of this study showed that the methanol extract of E. hirta plant had antibacterial activity against R. solanacearum, E. coli and S. aureus, thus can be used as a biocontrol agent of bacterial wilt disease in plants caused by R. solanacearum and human disease caused by E. coli and S. aureus. Keywords: Euphorbia hirta, Ralstonia solanacearum, Escherichia coli, Staphylococcus aureus, antibacterial activity, diffusion method