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Recovery of Sericin Protein from Silk Processing Wastewaters by Membrane Technology a Thesis Submitted to the Graduate School O RECOVERY OF SERICIN PROTEIN FROM SILK PROCESSING WASTEWATERS BY MEMBRANE TECHNOLOGY A THESIS SUBMITTED TO THE GRADUATE SCHOOL OF NATURAL AND APPLIED SCIENCES OF MIDDLE EAST TECHNICAL UNIVERSITY BY SAN İYE SEYLAN AYGÜN IN PARTIAL FULFILLMENT OF THE REQUIREMENTS FOR THE DEGREE OF MASTER OF SCIENCE IN ENGINEERING SCIENCES JULY 2008 Approval of the thesis: RECOVERY OF SERICIN PROTEIN FROM SILK PROCESSING WASTEWATERS BY MEMBRANE TECHNOLOGY submitted by SAN İYE SEYLAN AYGÜN in partial fulfillment of the requirements for the degree of Master of Science in Engineering Sciences Department, Middle East Technical University by, Prof. Dr. Canan Özgen Dean, Graduate School of Natural and Applied Sciences ─────────── Prof. Dr. Turgut Tokdemir Head of Department, Engineering Sciences ─────────── Prof. Dr. M. Ru şen Geçit Supervisor, Engineering Sciences Dept., METU ─────────── Dr. Gök şen Çapar Co- Supervisor, Engineering Sciences Dept., METU ─────────── Examining Committee Members Prof. Dr. Ülkü Yeti ş ─────────── Environmental Engineering Dept., METU Prof.Dr. M. Ru şen Geçit ─────────── Engineering Sciences Dept., METU Prof. Dr. Levent Yılmaz ─────────── Chemical Engineering Dept., METU Dr. Gök şen Çapar ────────── ─ Engineering Sciences Dept., METU Assist. Prof. Dr. Dilek Keskin ─────────── Engineering Sciences Dept., METU Date: 30.07.2008 I hereby declare that all information in this document has been obtained and presented in accordance with academic rules and ethical conduct. I also declare that, as required by these rules and conduct, I have fully cited and referenced all material and results that are not original to this work. Name, Last name : Signature : iii ABSTRACT RECOVERY OF SERICIN PROTEIN FROM SILK PROCESSING WASTEWATERS BY MEMBRANE TECHNOLOGY Aygün, Saniye Seylan M.S., Department of Engineering Sciences Supervisor : Prof. Dr. M. Ru şen Geçit Co-Supervisor : Dr. Gök şen Çapar July 2008, 182 pages Cocoon cooking wastewaters (CW) and silk degumming wastewaters (SDW) of silk processing industry were treated by membrane processes for sericin recovery. CW contains only sericin while SDW contains both sericin and soap. Sericin in CW had four molecular weight (MW) fractions; 175-200 kDa (Sericin-1), 70-90 kDa (Sericin- 2), 30-40 kDa (Sericin-3) and 10-25 kDa (Sericin-4). Two alternative process trains were developed for CW; 1. centrifugation + microfiltration + nanofiltration + precipitation, 2. centrifugation + microfiltration + nanofiltration + dialysis + precipitation. In the first process, a sericin/silkworm protein mixture was obtained with a sericin content of 39-46%. In the second one, however, a pure sericin product was obtained. The sericin recovery efficiency of the developed process train was found as 76%. Severe flux declines of 70-75% were observed in NF stage in both process trains. However, cleaning with 0.5 M NaOH and 190-200 mg/L free chlorine restored the fluxes by 83-127%. iv The MW of sericin in SDW was 110-120 kDa. The soap and sericin were separated in the pre-treatment stage consisting of centrifugation (pH 3.5, 10 min) and gravity settling (4 oC, 24 h). The ultrafiltration membrane with molecular weight cut-off of 5 kDa achieved 59% sericin recovery at pH 3.5, accompanied by severe flux decline of 88%. Furthermore, clean water flux was restored by only 31% via chemical cleaning. Keywords: Nanofiltration; Recovery; Sericin; Silk processing wastewater; Ultrafiltration v ÖZ İPEK İŞ LEME ATIKSULARINDAN MEMBRAN TEKNOLOJ İSİ İLE SER İSİN PROTE İNİNİN GER İ KAZANILMASI Aygün, Saniye Seylan Yüksek Lisans, Mühendislik Bilimleri Bölümü Tez Yöneticisi : Prof. Dr. M. Ru şen Geçit Ortak Tez Yöneticisi : Dr. Gök şen Çapar Temmuz 2008, 182 sayfa İpek endüstrisinin koza pi şirme (KA) ve ipek yumu şatma atıksuları ( İYA), serisin geri kazanımı için membran prosesleri ile arıtılmı ştır. İYA, serisin ve sabun içerirken; KA sadece serisin içermektedir. Koza pi şirme atıksuyunda dört ayrı molekül a ğırlı ğına sahip serisin bulunmu ştur; 175-200 kDa (Serisin-1), 70-90 kDa (Serisin-2), 30-40 kDa (Serisin-3) ve 10-25 kDa (Serisin-4). Koza atıksuları için iki alternatif süreç geli ştirilmi ştir; 1. santrifüj + mikrofiltrasyon + nanofiltrasyon + çökeltme, 2. santrifüj + mikrofiltrasyon + nanofiltrasyon + diyaliz + çökeltme. İlk süreçte, %39-46 serisin içeri ğine sahip olan serisin/ipekböce ği proteini karı şımı elde edilmi ştir. İkincisinde, ise saf serisin elde edilmi ştir. Geli ştirilen geri kazanım sürecinin serisini geri vi kazanma verimi %76 olarak bulunmu ştur. NF a şamasında her iki süreçte %70-75’lik ciddi akı azalmaları gözlenmi ştir. Ancak, 0,5 M NaOH ve 190-200 mg/L serbest klorla yapılan yıkama ile akılar %83-127 oranında iyile ştirilmi ştir. İYA’daki serisinin molekül a ğırlı ğı 110-120 kDa olarak bulunmu ştur. Atıksudaki sabun ve serisin, santrifüj (pH 3,5, 10 dk) ve cazibe ile çökmeyi (4 oC, 24 s) içeren ön-arıtma a şamasında ayrılmı ştır. Molekül a ğırlık ayırma sınırı 5 kDa olan ultrafiltrasyon membranı, %88’lik ciddi bir akı azalması ile birlikte, pH 3,5’te %59 oranında serisin geri kazanımı sa ğlamı ştır. Bunun yanı sıra, saf su akısı kimyasal yıkama ile sadece %31 oranında iyile ştirilebilmi ştir. Anahtar Sözcükler: Nanofiltrasyon; Geri kazanım; Serisin; İpek i şleme atıksuyu; Ultrafiltrasyon; vii To My Father, Secaattin Aygün... viii ACKNOWLEDGMENTS I would like to express my deepest gratitude to my supervisor Prof. Dr. M. Ru şen Geçit and co-supervisor Dr. Gök şen Çapar for their guidance, continuous support, encouragement, advice and trust during this study. I would like to thank Prof. Dr. Ülkü Yeti ş and Prof. Dr. Levent Yılmaz for their constructive suggestions and helpful discussions. I would like to present my special thank to Assist. Prof. Dr. Dilek Keskin for encouraging me for this master. Also, I would like to thank Assist. Prof. Dr. Ay şen Tezcaner for her helpful suggestions and discussions. I would like to thank the Environmental Engineering and Chemical Engineering Department staff, especially Gizem U ğurlu and Nimet Uzal for their help about laboratory equipments. Also, I would like to mention Baki Akda ğ and Turgut Sipahio ğlu for providing samples and information about silk processing wastewaters. TÜB İTAK is gratefully acknowledged for the financial support via project number 106M062. Many thanks to my friends, Aslı Erdo ğ, Ay şegül Kavas, Burçin Başar, Ömer Aktürk, Özge Erdemli and Özlem Aydın for their friendship and continuous support throughout this research. My sincer thanks go to my mother, Semra Aygün; my brother, Taylan Aygün and his wife, Ye şim Aygün for their endless support and encouragement during this study. And, I wish to thank my little nephew, Ata Yi ğit Aygün for amusing me with his joy and sweetness. Finally, special thanks to my dear fiance, Memduh Karalar for beautifying my life with his endless support and endless love. ix TABLE OF CONTENTS ABSTRACT...................................................................................................... iv ÖZ.................................................................................................................... vi DEDICATION................................................................................................... viii ACKNOWLEDGMENTS................................................................................... ix TABLE OF CONTENTS................................................................................... x LIST OF TABLES............................................................................................. xiv LIST OF FIGURES........................................................................................... xvi ABBREVIATIONS............................................................................................ xix CHAPTER 1. INTRODUCTION................................................................................ 1 1.1. General Information...................................................................... 1 1.2. Aim and Scope of the Study......................................................... 3 2. SILK PROCESSING WASTEWATERS............................................. 8 2.1. Sericulture................................................................................... 8 2.2. Silk Processing............................................................................ 9 2.3. Characteristics of Silk Processing Wastewaters......................... 11 2.4. Treatment of Silk Processing Wastewaters................................. 15 3. MEMBRANE PROCESSES............................................................... 19 3.1. Historical Background.................................................................. 19 3.2. Definitions.................................................................................... 20 3.3. Pressure-Driven Membrane Processes...................................... 22 3.3.1. Microfiltration..................................................................... 23 3.3.2. Ultrafiltration...................................................................... 24 3.3.3. Nanofiltration..................................................................... 25 3.3.4. Reverse Osmosis.............................................................. 25 x 3.4. Membrane Fouling and Cleaning................................................ 26 4. LITERATURE REVIEW ON SERICIN RECOVERY FROM SILK PROCESSING WASTEWATERS..................................................... 28 4.1. Characteristics of Sericin...........................................................
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