جداسازی و شناسایی مولکولی باکتری های تولیدکننده کاروتنوئید

نویسندگان

1 گروه زیست شناسی، دانشکده علوم پایه، دانشگاه آزاد اسلامی، واحد علوم و تحقیقات، تهران، ایران.

2 گروه میکروب‌شناسی و انگل‌شناسی ، دانشکدۀ پزشکی، دانشگاه علوم پزشکی بوشهر، بوشهر، ایران.

3 دانشگاه شهرکرد

4 گروه میکروب‌شناسی، دانشکدۀ پزشکی، دانشگاه علوم پزشکی گلستان، گرگان، ایران.

چکیده
تولید رنگدانه از باکتری­ ها به علت مقرون به صرفه بودن، محصول بیشتر و استخراج آسان­تر نسبت به سایر منابع از اهمیت خاصی برخوردار است. کاروتنوئید­ها، از مهمترین رنگدانه­ ها با خواص آنتی ­اکسیدانی، پیش ­ساز ویتامینA ، افزایش ­دهنده تولید آنتی ­بادی، ضد تومور و پیشگیری کننده از بیماری ­های قلبی-عروقی هستند. هدف از این مطالعه جداسازی و شناسایی مولکولی باکتری­ های تولیدکنندۀ رنگدانه­ کاروتنوئیدی و آنالیز آن با روش HPLC بود. تعداد 20 نمونه خاک از مناطق مختلف شهر تهران جمع ­آوری شد. پس از رقت­ سازی، نمونه ­ها بر روی محیطBHI آگار کشت و در دمای 37 درجه­ سانتی­ گراد گرماگذاری شد. باکتری­ های تولیدکننده رنگدانه برای شناسایی بیشتر انتخاب و استخراج رنگدانۀ آنها بوسیله­ متانول انجام شد. غربالگری باکتری­ های تولیدکننده­ رنگدانه در دو سطح انجام گرفت: انتخاب سویه­ ها با استفاده از رنگ قابل مشاهده؛ آنالیز عصاره ها با طیف سنجی UV-VIS و تأیید توسط کروماتوگرافی مایع با کارایی بالا .(HPLC) ابتدا، جدایه ­ها با استفاده از روش­ های فنوتیپی شناسایی شد و ژن 16S rDNA آنها با روش PCR تکثیر و تعیین توالی شد. Staphylococcus epidermidis، Micrococcus aloeverae، Citricoccus alkalitolerans، Rhodococcus zopfii، Arthrobacter agilis،Dietzia natronolimnaea وRhodococcus ruber به­ عنوان سویه های تولیدکنندۀ کاروتنوئید شناسایی شدند .بالاترین میزان جذب با استفاده از آنالیز طیف ­سنجی UV-VIS در Staphylococcus epidermidis و Dietzia natronolimnaea مشاهده شد. آنالیز HPLC نشان داد که کاروتنوئیدهای تولید شده در مقایسه با منحنی بتا­کاروتن استاندارد، متعلق به بتاکاروتن هستند. میکروارگانیسم­ ها منبع بالقوه ­ای برای تولید کارتنوئیدها هستند. در این مطالعه، ما دو جنس باکتری (Staphylococcus epidermidis و Dietzia natronolimnaea) با توانایی بالای تولید کارتنوئید معرفی کردیم.

کلیدواژه‌ها


عنوان مقاله English

Isolation and molecular identification of carotenoid-producing bacteria

نویسندگان English

Razieh sadat Solouki Nezhad 1
Hanieh Asaadi 2
Yaser Eshaghi Milasi 3
Sajjad Yazdansetad 4
1 Department of Biology, Faculty of Basic Sciences, Islamic Azad University, Science and Research Branch, Tehran, Iran.
2 Department of Microbiology and Parasitology, Faculty of Medicine, Bushehr University of Medical Sciences, Bushehr, Iran.
3 Shahrekord University
4 Department of Microbiology, School of Medicine, Golestan University of Medical Sciences, Gorgan, Iran.
چکیده English

The production of pigments from bacteria is significant due to the low cost, high yield and ease of extract compared with other sources. Carotenoids are one of the most important pigments with antioxidant properties which are the precursor of vitamin A synthesis and have antibody overproduction ability, anti-tumor activity and inhibitory effect on the cardiovascular disease. The present study aimed to isolate and identify carotenoid-producing bacteria by high-performance liquid chromatography (HPLC) analysis of their carotenoid pigments. Twenty soil samples were collected from different regions of Tehran. After serial dilution each sample was cultured on BHI agar medium and incubated at 37°C. The pigment-producing bacteria were selected for further identification and their pigments were extracted by methanol. The screening was carried out at two levels: i) selection of the strains by visual color inspection, ii) analysis of the pigment extracts by UV-VIS spectroscopy and HPLC. The isolates were identified by phenotypic methods and their 16S rDNA gene was amplified by PCR method and sequenced. Staphylococcus epidermidis, Micrococcus aloeverae, Citricoccus alkalitolerans, Rhodococcus zopfii, Arthrobacter agilis, Dietzia natronolimnaea and Rhodococcus ruber were identified as carotenoid-producing strains. The highest rate of absorption was observed using UV-VIS analysis in Staphylococcus epidermidis and Dietzia natronolimnaea. The comparison of HPLC analysis with the standard β-carotene curve showed that the carotenoids were beta-carotene. Micro-organisms are a potential source in the production of pigments. In this study we introduced two genera of bacteria (Staphylococcus epidermidis and Dietzia natronolimnaea) with carotenoid-producing ability.


کلیدواژه‌ها English

β-carotene
carotenoids
chromatography
pigment
spectrophotometry
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  • تاریخ دریافت 18 خرداد 1405
  • تاریخ انتشار 18 خرداد 1405