Isolation and Purifi cation of Chitinase Bacillus sp. D2 Isolated from Potato Rhizosfer

• Main Authors: Sebastian Margino, Chatarina Behar, Widya Asmara
• Format: Article
• Language: English
• Published: Universitas Gadjah Mada, Yogyakarta 2015-11-01
• Series: Indonesian Journal of Biotechnology
• Online Access: http://journal.ugm.ac.id/ijbiotech/article/view/7851

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table.MsoNormalTable {mso-style-name:"Table Normal"; mso-tstyle-rowband-size:0; mso-tstyle-colband-size:0; mso-style-noshow:yes; mso-style-priority:99; mso-style-qformat:yes; mso-style-parent:""; mso-padding-alt:0cm 5.4pt 0cm 5.4pt; mso-para-margin:0cm; mso-para-margin-bottom:.0001pt; mso-pagination:widow-orphan; font-size:11.0pt; font-family:"Calibri","sans-serif"; mso-ascii-font-family:Calibri; mso-ascii-theme-font:minor-latin; mso-fareast-font-family:"Times New Roman"; mso-fareast-theme-font:minor-fareast; mso-hansi-font-family:Calibri; mso-hansi-theme-font:minor-latin; mso-bidi-font-family:"Times New Roman"; mso-bidi-theme-font:minor-bidi;} --> <!--[endif] --> <p class="MsoNormal"><span style="font-size: 9.5pt; font-family: &quot;BookAntiqua&quot;,&quot;serif&quot;;" lang="EN-GB">Potato Cyst Nematodes (</span><em><span style="font-size: 9.5pt; font-family: &quot;BookAntiqua-Italic&quot;,&quot;serif&quot;;" lang="EN-GB">Globodera rostochiensis) </span></em><span style="font-size: 9.5pt; font-family: &quot;BookAntiqua&quot;,&quot;serif&quot;;" lang="EN-GB">is one of the important potato&rsquo;s pests and caused economic</span></p> <p class="MsoNormal"><span style="font-size: 9.5pt; font-family: &quot;BookAntiqua&quot;,&quot;serif&quot;;" lang="EN-GB">looses up to 70% in the several centrals of potato plantations in Indonesia. Potato Cyst Nematodes (PCN) shell</span></p> <p class="MsoNormal"><span style="font-size: 9.5pt; font-family: &quot;BookAntiqua&quot;,&quot;serif&quot;;" lang="EN-GB">component of egg shell containing chitin (inner layer) and vitelline/protein (outer layer), so the purpose of</span></p> <p class="MsoNormal"><span style="font-size: 9.5pt; font-family: &quot;BookAntiqua&quot;,&quot;serif&quot;;" lang="EN-GB">research was to fi nd out of chitin degrading bacteria for controlling of egg&rsquo;s PCN by cutting of their life cycle.</span></p> <p class="MsoNormal"><span style="font-size: 9.5pt; font-family: &quot;BookAntiqua&quot;,&quot;serif&quot;;" lang="EN-GB">The results showed that </span><em><span style="font-size: 9.5pt; font-family: &quot;BookAntiqua-Italic&quot;,&quot;serif&quot;;" lang="EN-GB">Bacillus </span></em><span style="font-size: 9.5pt; font-family: &quot;BookAntiqua&quot;,&quot;serif&quot;;" lang="EN-GB">sp. D2 isolated from potato rhizosphere could produce extra cellular chitinase</span></p> <p class="MsoNormal"><span style="font-size: 9.5pt; font-family: &quot;BookAntiqua&quot;,&quot;serif&quot;;" lang="EN-GB">in the medium containing of 0.20% colloidal chitin and fermented for 72 hours. Result of chitinase purifi cation</span></p> <p class="MsoNormal"><span style="font-size: 9.5pt; font-family: &quot;BookAntiqua&quot;,&quot;serif&quot;;" lang="EN-GB">using ammonium sulphate precipitation and </span><em><span style="font-size: 9.5pt; font-family: &quot;BookAntiqua-Italic&quot;,&quot;serif&quot;;" lang="EN-GB">DEAE-Cellulose </span></em><span style="font-size: 9.5pt; font-family: &quot;BookAntiqua&quot;,&quot;serif&quot;;" lang="EN-GB">ion-exchange chromatography showed a specifi c</span></p> <p class="MsoNormal"><span style="font-size: 9.5pt; font-family: &quot;BookAntiqua&quot;,&quot;serif&quot;;" lang="EN-GB">activity 2691,052 U/mg and analyzing using SDS-PAGE 12.5% resulted in molecular weight 30 kDa. The</span></p> <p class="MsoNormal"><span style="font-size: 9.5pt; font-family: &quot;BookAntiqua&quot;,&quot;serif&quot;;" lang="EN-GB">apparent K</span><span style="font-size: 5.5pt; font-family: &quot;BookAntiqua&quot;,&quot;serif&quot;;" lang="EN-GB">m </span><span style="font-size: 9.5pt; font-family: &quot;BookAntiqua&quot;,&quot;serif&quot;;" lang="EN-GB">and V</span><span style="font-size: 5.5pt; font-family: &quot;BookAntiqua&quot;,&quot;serif&quot;;" lang="EN-GB">max </span><span style="font-size: 9.5pt; font-family: &quot;BookAntiqua&quot;,&quot;serif&quot;;" lang="EN-GB">of chitinase towards colloidal chitin were 2 mg/ml and 2.2 &mu;g/h, respectively.</span></p> <p class="MsoNormal"><span style="font-size: 9.5pt; font-family: &quot;BookAntiqua&quot;,&quot;serif&quot;;" lang="EN-GB">&nbsp;</span></p>