جداسازی، شناسایی و تعیین ویژگی دو گونه تریکوسپورون مخمری تجزیه کننده فنل جداسازی شده از پساب کارخانه کک‌سازی زرند کرمان

نوع مقاله : مقاله پژوهشی

نویسندگان

1 دانشیار میکروبیولوژی، گروه زیست‌شناسی، دانشکده علوم، دانشگاه شهید باهنر کرمان، ایران

2 الف: دانش‌آموخته کارشناسی ارشد میکروبیولوژی، گروه میکروبیولوژی، واحد کرمان، دانشگاه آزاد اسلامی، کرمان، ایران ب: دانش‌آموخته کارشناسی ارشد، گروه میکروبیولوژی، واحد علوم تحقیقات کرمان، دانشگاه آزاد اسلامی، کرمان، ایران

3 دانشجوی دکترای میکروبیولوژی، عضو هیئت علمی واحد فلاورجان، دانشگاه آزاد اسلامی، اصفهان، ایران

چکیده

فنل و ترکیبات فنلی، آلوده‌کننده‌های محیطی هستند که در پساب صنایع مختلف از جمله زغال سنگ، پالایشگاه‌های نفت، کارخانه‌های فراورده‌های شیمیایی و پتروشیمی وجود دارند. حذف فنل از خروجی پساب صنایع اهمیت زیادی در حفظ محیط زیست دارد. تجزیه فنل، معمولاً به‌وسیله روش‌های فیزیکوشیمیایی صورت می‌گیرد، اما این روش‌ها هزینه‌بر بوده و واسطه‌های خطرناک تولید می‌کنند.تجزیه بیولوژیکی فنل در سال‌های اخیر مورد توجه قرار گرفته است. از بین میکروارگانیسم‌ها، مخمرها اهمیت خاصی در تجزیه فنل دارند. در این پژوهش، مخمرهای تجزیه‌کننده فنل ابتدا از نمونه‌های محیطی (خاک و پساب ) کارخانه کک‌سازی زرند کرمان جداسازی شد. سپس کل مخمرهای هتروتروف و تجزیه‌کننده شمارش شدند که نمونه‌های خاک، دارای میزان بالاتری از مخمرهای تجزیه‌کننده نسبت به نمونه‌های پساب بودند. پس از سه بار پاساژ دادن مخمرها، میزان رشد آنها اندازه‌گیری شد که مخمرهای K1 و K11 دارای بیشترین میزان رشد بودند. همچنین همین مخمرها قادر به حذف فنل بودند که سنجش آن توسط معرف گیبس صورت گرفت. سپس اثر چهار غلظت مختلف فنل (05/0، 125/0، 2/0 و 275/0 گرم در لیتر) بررسی شد و دو الگوی تجزیه‌ای در این مخمرها مشاهده شد. همچنین مقدار فعالیت امولسیون‌کنندگی و هیدروفوبیسیته هر یازده مخمر اندازه‌گیری شد. در نهایت پس از یک‌سری آزمون‌های بیوشیمیایی، شناسایی مولکولی مخمرهای قوی در تجزیه فنل با تکثیر قسمتی از ناحیه ژنی 18S rRNA با پرایمرهای ویژه این ژن (Euk-A, Euk-B) انجام شد. توالی حاصله علیه بانک‌های ژنی مقایسه شد و بالاترین همولوژی ( بالاتر از 98 درصد) به‌عنوان جنس و گونه مخمر تجزیه‌کننده تعیین شد. مخمرهای جداسازی شده مربوط به جنس‌های تریکوسپورون مونتاویدینس و تریکوسپورون کوتانئوم  بودند.

کلیدواژه‌ها

موضوعات


عنوان مقاله [English]

Isolation, Identification and Characterization of Two Phenol-Degrading Species of Trichosporon Isolated from Wastewater of Zarand Coking Plant -Kerman

نویسندگان [English]

  • Mehdi Hassanshahian 1
  • Maryam Karimi 2
  • Zarrindokht Emami 3
1 Assoc. Prof. of Microbiology, Department of Biology, Faculty of Sciences, Shahid Bahonar University, Kerman, Iran
2
3 PhD Student of Microbiology, Faculty Member of Falavarjan Branch, Islamic Azad University, Isfaha, Iran
چکیده [English]

Phenol and phenolic compounds are environmental contaminants that are commonly found in industrial effluents from oil refineries, coal mines, and petrochemical plants. Phenol removal from industrial effluent is, therefore, of extreme importance for environmental protection. Phenol degradation is commonly accomplished by physicochemical methods that are not only costly but also produce hazardous metabolites. Recently, phenol biodegradation has attracted much attention due to its lower process cost. In this study, yeast, as a phenol-degrading agent, was initially isolated from samples in the surrounding environment (soil and water) taken from Zarand coking plant. The total heterotrophic and biodegrading yeasts were then counted. Compared to effluent samples, soil samples were found to exhibit higher yeast degrader counts. Yeast growths were measured after three passages and the two strains K1 and K11 were recorded as the ones with the highest growth rates. These same yeasts were the ones capable of removing phenol as evidenced by the remaining phenol content measured by the Gibbs reagent. The effects of four different phenol concentrations (0.1, 0.125, 0.2, and 0.275) were activities were measured in all the eleven yeasts isolated. Finally, part of the 18S rRNA gene section was cultured using primers (Euk-A, Euk-B) especially used for this gene and a series of biochemical tests were run for the molecular identification of the yeasts predominantly involved in phenol degradation. The sequences obtained were compared against the available gene banks and the highest homology (greater than 98%) was introduced as the genus and species of phenol degrader yeasts. The isolated yeasts belonged to the genera Trichosporon montevideense and Trichosporon cutaneum.  

کلیدواژه‌ها [English]

  • Biodegradation
  • Phenol
  • Coal Tar
  • Yeast
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