Mutagenic Improvement of Xylanase Production from Xylanolytic Bacteria and its Phylogenetic Analysis

Authors

  • CHUSNUL HANIM
  • LIES MIRA YUSIATI
  • MUHAMMAD NUR CAHYANTO
  • ALI WIBOWO

DOI:

https://doi.org/10.5454/mi.7.2.2

Keywords:

ethidium bromide, ethyl methanesulfonate, mutagenesis, xylanolytic bacteria

Abstract

This study was conducted to obtain xylanolytic mutants that have higher xylanase activity than their wildtype counterparts. A mutant with the best xylanolytic activity was selected and identified based on its 16S rRNA sequence. Its optimum growth condition was also characterized and its phylogenetic relations to other xylanolytic bacteria were analzsed. Wild type xylanolytic alkalophlic bacteria were grown in medium containing xylan as a substrate. Mutation was performed using ethidium bromide (EtBr) or ethyl methanesulfonate (EMS) at

concentrations 50, 100, and 150 mg mL-1 and times of exposure 30, 60, 90, and 120 min for each treatment. Twenty two mutants were obtained from EtBr and 24 mutants from EMS mutageneses. The mutants were analyzed for their capability to secrete xylanase into xylan medium containing xylose or glucose or glycerol. Growth optimizations of the mutant were done in media with pH range 6-11 and temperature range 30 to 60 °C. Mutant number 19, which was obtained by treatment using 50 mg mL-1 EMS for 120 min, had the highest xylanase activity (15.057 U g-1). This activity was obtained at optimum growth conditions: pH 9.5 and temperature 55 °C. Chromosomal DNA of this mutant was extracted and amplified by PCR using 16S rRNA gene specific primers. The amplified fragments were sequenced by dideoxynucleotide chain terminator method. The phylogenetic analysis based on 16S rRNA gene sequence showed that mutant 19 was closed to an anaerobic xylanase producing bacteria.

Downloads

Published

2013-07-08

How to Cite

HANIM, C., YUSIATI, L. M., CAHYANTO, M. N., & WIBOWO, A. (2013). Mutagenic Improvement of Xylanase Production from Xylanolytic Bacteria and its Phylogenetic Analysis. Microbiology Indonesia, 7(2), 2. https://doi.org/10.5454/mi.7.2.2

Issue

Section

Articles
                                                                  ADOConnection.CacheExecute(3600, SELECT 				o.submission_id, 				MAX(s.context_id) AS journal_id, 				MAX(i.date_published) AS i_pub, 				MAX(p.date_published) A..., Array[2]) % line  132, file: DAO.inc.php
                                                               DAO.retrieveCached(SELECT 				o.submission_id, 				MAX(s.context_id) AS journal_id, 				MAX(i.date_published) AS i_pub, 				MAX(p.date_published) A..., Array[2], 3600) % line   91, file: ArticleSearchDAO.inc.php
                                                            ArticleSearchDAO.getPhraseResults(Object:Journal, Array[1], null, null, , 500, 1) % line  201, file: SubmissionSearch.inc.php
                                                         SubmissionSearch._getMergedPhraseResults(Object:Journal, Array[1], , null, null, 500, 1) % line  146, file: SubmissionSearch.inc.php
                                                      SubmissionSearch._getMergedKeywordResults(Object:Journal, Array[4], , null, null, 500, 1) % line  194, file: SubmissionSearch.inc.php
                                                   SubmissionSearch._getMergedPhraseResults(Object:Journal, Array[4], null, null, null, 500, 1) % line  146, file: SubmissionSearch.inc.php
                                                SubmissionSearch._getMergedKeywordResults(Object:Journal, Array[3], null, null, null, 500, 1) % line  132, file: SubmissionSearch.inc.php
                                             SubmissionSearch._getMergedArray(Object:Journal, Array[1], null, null) % line  263, file: SubmissionSearch.inc.php
                                          SubmissionSearch.retrieveResults(Object:Request, Object:Journal, Array[1], null, null, null, Object:DBResultRange, Array[1]) % line   92, file: RecommendBySimilarityPlugin.inc.php
                                       RecommendBySimilarityPlugin.callbackTemplateArticlePageFooter(Templates::Article::Footer::PageFooter, Array[3]) % line  107, file: HookRegistry.inc.php
                                    HookRegistry.call(Templates::Article::Footer::PageFooter, Array[3]) % line 1256, file: PKPTemplateManager.inc.php
                                 PKPTemplateManager.smartyCallHook(Array[1], Object:Smarty_Internal_Template) % line   43, file: 42362804174f266c598cdb32c3208fc745b92101^1c372c95cd85572e0fbc9a53d0323a0b229cfc78_0.app.frontendpagesarticle.tpl.php
                              content_6082325297a527_48332634(Object:Smarty_Internal_Template) % line  123, file: smarty_template_resource_base.php
                           Smarty_Template_Resource_Base.getRenderedTemplateCode(Object:Smarty_Internal_Template) % line  114, file: smarty_template_compiled.php
                        Smarty_Template_Compiled.render(Object:Smarty_Internal_Template) % line  216, file: smarty_internal_template.php
                     Smarty_Internal_Template.render(false, 1) % line  232, file: smarty_internal_templatebase.php
                  Smarty_Internal_TemplateBase._execute(Object:Smarty_Internal_Template, null, 42362804174f266c598cdb32c3208fc745b92101, null, 1) % line  134, file: smarty_internal_templatebase.php
               Smarty_Internal_TemplateBase.display(frontend/pages/article.tpl, null, 42362804174f266c598cdb32c3208fc745b92101, null) % line  924, file: PKPTemplateManager.inc.php
            PKPTemplateManager.display(frontend/pages/article.tpl) % line  287, file: ArticleHandler.inc.php
         ArticleHandler.view(Array[1], Object:Request) % line  391, file: PKPRouter.inc.php
      PKPRouter._authorizeInitializeAndCallRequest(Array[2], Object:Request, Array[1], false) % line  231, file: PKPPageRouter.inc.php
   PKPPageRouter.route(Object:Request) % line  143, file: Dispatcher.inc.php
Dispatcher.dispatch(Object:Request) % line  281, file: PKPApplication.inc.php
PKPApplication.execute() % line   68, file: index.php