Luận văn Nghiên cứu quy trình cố định nấm men trên chất màng BC và ứng dụng và quá trình lên men sản xuất rượu vang

Chương 1: GIỚI THIỆU Như chúng ta đã biết, rượu vang là một trong những sản phẩm lên men có lịch sử lâu đời nhất. Theo các tài liệu cổ, rượu vang có xuất xứ từ các nước Ả Rập vào khoảng thế kỷ 18 hay 19 trước Công nguyên và cho đến nay, rượu vang đã trở nên phổ biến khắp nơi trên thế giới. Rượu vang được ưa thích trước hết là do hương vị hết sức đặc trưng mà không loại sản phẩm nào có thể thay thế, sau đó là do nó đem lại cảm giác sảng khoái và sức khỏe cho người uống. Đầu tiên, rượu vang được tạo ra bằng quá trình lên men tự phát. Sau đó, người ta đã tiến hành phân lập các chủng nấm men và cấy vào dịch lên men loài Saccharomyces cerevisiae thuần thiết để thúc đẩy quá trình lên men và tăng chất lượng của rượu vang. Tuy nhiên, việc sử dụng nấm men tự do để lên men có khá nhiều nhược điểm: - Thời gian lên men dài, năng suất lên men thấp. - Tiêu tốn nhiều năng lượng để tách nấm men ra khỏi rượu vang sau quá trình lên men. - Khả năng tái sử dụng nấm men rất thấp. - Khó tự động hóa quá trình lên men. Để khắc phục các nhược điểm này, nhiều nghiên cứu về việc sử dụng nấm men cố định trong sản xuất rượu vang đã được thực hiện. Các nghiên cứu này đều cho thấy nấm men cố định có nhiều ưu điểm hơn hẳn so với nấm men tự do: - Tăng tốc độ sử dụng cơ chất, rút ngắn thời gian lên men. - Ổn định hoạt tính của nấm men. Các chất mang cố định có tác dụng như là một tác nhân bảo vệ tế bào chống lại những ảnh hưởng bất lợi của pH, nhiệt độ, dung môi và ngay cả các kim loại nặng. - Dễ dàng tách nấm men ra khỏi sản phẩm sau quá trình lên men, do đó làm giảm chi phí về thiết bị và năng lượng. - Có thể tái sử dụng nấm men nhiều lần. - Tạo điều kiện thuận lợi cho việc tối ưu hóa và tự động hóa quá trình lên men. Các nghiên cứu trước đây về ứng dụng nấm men cố định trong sản xuất rượu vang chủ yếu khảo sát quá trình sử dụng đường, quá trình sinh tổng hợp cồn và sự hình thành các hợp chất hương mà ít đề cập đến động học quá trình sinh trưởng của nấm men, quá trình sử dụng các hợp chất nitơ cũng như sự chuyển hóa của các acid hữu cơ trong suốt quá trình lên men chính. Chúng tôi cũng tìm thấy rất ít các nghiên cứu về ảnh hưởng của một số yếu tố công nghệ trong quá trình sản xuất đến động học quá trình lên men rượu vang sử dụng nấm men cố định như ảnh hưởng của hàm lượng tannin, hàm lượng SO2 trong dịch nho ban đầu. Tuy nhiên, các nghiên cứu này cũng chỉ khảo sát quá trình sinh tổng hợp cồn mà chưa khảo sát một cách toàn diện ảnh hưởng của các yếu tố này đến tất cả các mặt trong quá trình lên men chính. Trên cơ sở đó, chúng tôi đề xuất đề tài nghiên cứu “Khảo sát ảnh hưởng của các yếu tố công nghệ đến động học quá trình lên men rượu vang nho sử dụng nấm men cố định”. Chúng tôi tiến hành khảo sát ảnh hưởng của 4 yếu tố công nghệ, bao gồm: pH, hàm lượng đường, hàm lượng SO2 và hàm lượng tannin ban đầu. Tuy nhiên, trong phạm vi bài luận văn này, tôi chỉ xin trình bày sư ảnh hưởng của 2 yếu tố là hàm lượng đường và hàm lượng tannin ban đầu. Kết quả về sự ảnh hưởng của pH và hàm lượng SO2 trong dịch nho đến quá trình lên men sẽ do một bạn khác trình bày. Trong nghiên cứu này, chúng tôi sử dụng chất mang là alginate - một polysaccharide thu được từ rong mơ phân bố dọc theo bờ biển miền Trung của Việt Nam. Chúng tôi lần lượt khảo sát ảnh hưởng của hàm lượng đường và hàm lượng tannin ban đầu đến các vấn đề sau: - Động học quá trình sinh trưởng của nấm men. - Động học quá trình sử dụng cơ chất: đường khử, nitơ amin và nitơ vô vơ. - Động học quá trình hình thành sản phẩm: do thời gian hạn hẹp và chưa có điều kiện về kinh phí nên trong phạm vi bài luận văn này, chúng tôi chỉ trình bày sản phẩm quan trọng nhất của quá trình lên men là cồn mà chưa trình bày các hợp chất hương. Và chúng tôi cũng khảo sát thêm sự chuyển hóa của các acid hữu cơ trong suốt quá trình lên men chính.

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rung bình  ñoä leäch chuaån cuûa 3 maãu ñoäc laäp. Caùc giaù trò coù kyù hieäu khaùc nhau bieåu thò söï khaùc nhau coù nghóa (P < 0,05). Quaù trình chuyeån hoùa caùc acid höõu cô Quaù trình chuyeån hoùa caùc acid höõu cô cuõng ñöôïc chuùng toâi ñaùnh giaù thoâng qua söï thay ñoåi pH, haøm löôïng acid toång vaø haøm löôïng acid deã bay hôi nhö ôû phaàn 4.1.3.2.  LINK Excel.Chart.8 "F:\\Tai Lieu hoc tap\\Tu lieu\\Ninh beo\\KET QUA THI NGHIEM\\Ket qua da xu ly xong\\sinh truong tu do lon hon co dinh\\Tannin-chung.xls" "do thi pH" \a \p Error! Not a valid link. Hình  STYLEREF 1 \s 4. SEQ Hình \* ARABIC \s 1 33: Söï thay ñoåi pH trong quaù trình leân men röôïu vang, söû duïng naám men töï do vaø naám men coá ñònh trong gel alginate (haøm löôïng tannin ban ñaàu thay ñoåi trong khoaûng 1,8 – 17,8g/L). Keát quaû khaûo saùt giaù trò pH cuûa chuùng toâi cho thaáy raèng pH vaø haøm löôïng acid toång cuûa dòch leân men baèng naám men coá ñònh luoân thaáp hôn pH cuûa dòch leân men baèng naám men töï do (hình 4.33 vaø hình 4.34). Ñieàu naøy ñaõ ñöôïc giaûi thích ôû phaàn 4.1.3.2. Beân caïnh ñoù, chuùng toâi cuõng thaáy raèng khi haøm löôïng tannin taêng thì pH taêng vaø haøm löôïng acid toång cuõng taêng. Hieän töôïng naøy coù theå ñöôïc giaûi thích laø do, khi haøm löôïng tannin taêng, khaû naêng khueách taùn cô chaát vaøo beân trong teá baøo naám men bò giaûm, toác ñoä leân men chaäm, neân ñaõ taïo ñieàu kieän cho caùc loaïi vi khuaån chuyeån hoùa ñöôøng thaønh acid acetic vaø acid lactic cuõng nhö caùc loaïi vi khuaån chuyeån hoùa caùc acid maïnh hôn nhö acid citric, acid tartaric vaø acid malic thaønh caùc acid yeáu hôn neân ñaõ laøm taêng haøm löôïng acid toång, ñoàng thôøi laïi laøm taêng pH dòch leân men.  LINK Excel.Chart.8 "F:\\Tai Lieu hoc tap\\Tu lieu\\Ninh beo\\KET QUA THI NGHIEM\\Ket qua da xu ly xong\\So lieu da xu ly-Luong\\Tannin - Luong.xls" "Do thi acid tong" \a \p Error! Not a valid link. Hình  STYLEREF 1 \s 4. SEQ Hình \* ARABIC \s 1 34: Söï thay ñoåi haøm löôïng acid toång trong quaù trình leân men röôïu vang söû duïng naám men töï do vaø naám men coá ñònh trong gel alginate (haøm löôïng tannin ban ñaàu thay ñoåi trong khoaûng 1,8 – 17,8g/L). Cuõng töông töï nhö vaäy, haøm löôïng acid deã bay hôi cuûa naám men coá ñònh thaáp hôn so vôùi naám men töï do vaø nhìn chung, khi haøm löôïng tannin taêng thì haøm löôïng acid deã bay hôi cuõng taêng (hình 4.35). So vôùi khi khaûo saùt aûnh höôûng cuûa haøm löôïng ñöôøng, chuùng toâi nhaän thaáy raèng, giaù trò pH cuûa caùc maãu tannin phaân taùn nhieàu hôn (hình 4.15 vaø hình 4.33). Ñieàu ñoù coù nghóa laø tannin aûnh höôûng raát lôùn ñeán giaù trò pH cuûa maãu, coøn ñöôøng thì ít aûnh höôûng hôn. Khi haøm löôïng tannin caøng taêng, thôøi gian ñeå pH ñaït ñeán giaù trò cöïc tieåu caøng daøi. Hay noùi caùch khaùc, quaù trình toång hôïp acid succinic dieãn ra chaäm hôn. Nhö ñaõ ñeà caäp ôû phaàn 4.1.3.2, acid succinic laø saûn phaåm cuûa quaù trình sinh tröôûng cuûa naám men, cho neân nhöõng hieän töôïng naøy coù theå deã daøng ñöôïc giaûi thích laø do tannin öùc cheá maïnh quaù trình sinh tröôûng cuûa naám men hôn laø aâ3 Toùm laïi, qua vieäc khaûo saùt söï chuyeån hoùa cuûa caùc acid höõu cô khi haøm löôïng tannin ban ñaàu thay ñoåi, chuùng toâi cuõng nhaän thaáy raèng naám men coá ñònh coù giaù trò pH, haøm löôïng acid toång vaø haøm löôïng acid deã bay hôi thaáp hôn so vôùi naám men töï do. Nhôø ñoù, ñoä beàn sinh hoïc, hoùa lyù vaø chaát löôïng caûm quan cuûa saûn phaåm leân men baèng naám men coá ñònh cuõng toát hôn so vôùi naám men töï do.  LINK Excel.Chart.8 "F:\\Tai Lieu hoc tap\\Tu lieu\\Ninh beo\\KET QUA THI NGHIEM\\Ket qua da xu ly xong\\So lieu da xu ly-Luong\\Tannin - Luong.xls" "Do thi acid de bay hoi" \a \p Error! Not a valid link. Hình  STYLEREF 1 \s 4. SEQ Hình \* ARABIC \s 1 35: Söï thay ñoåi haøm löôïng acid deã bay hôi trong quaù trình leân men röôïu vang, söû duïng naám men töï do vaø naám men coá ñònh trong gel alginate (haøm löôïng tannin ban ñaàu thay ñoåi trong khoaûng 1,8 – 17,8g/L). Keát luaän chung Qua keát quaû khaûo saùt aûnh höôûng cuûa haøm löôïng tannin ñeán ñoäng hoïc quaù trình leân men röôïu vang söû duïng naám men coá ñònh trong gel alginate, chuùng toâi coù moät soá keát luaän sau: Khi haøm löôïng tannin thaáp (1,8 – 2,8g/L) thì maät ñoä teá baøo cöïc ñaïi vaø toác ñoä sinh tröôûng rieâng cöïc ñaïi cuûa teá baøo coá ñònh thaáp hôn so vôùi teá baøo töï do. Nhöng khi haøm löôïng tannin cao thì maät ñoä teá baøo cöïc ñaïi vaø toác ñoä sinh tröôûng rieâng cöïc ñaïi cuûa teá baøo coá ñònh laïi cao hôn so vôùi teá baøo töï do. Nhö vaäy, khaû naêng sinh tröôûng cuûa naám men coá ñònh ít bò aûnh höôûng bôûi haøm löôïng tannin hôn so vôùi naám men töï do. Toác ñoä söû duïng ñöôøng cuõng nhö toác ñoä sinh toång hôïp coàn cuûa naám men coá ñònh cao hôn so vôùi naám men töï do nhöng hieäu suaát sinh toång hôïp coàn thì khoâng cao hôn. Khaû naêng söû duïng nitô ammonium cuõng nhö nitô höõu cô cuûa teá baøo coá ñònh cao hôn so vôùi teá baøo töï do. pH vaø acid toång cuûa dòch leân men baèng naám men coá ñònh thaáp hôn, do ñoù laøm taêng ñoä oån ñònh veà maët sinh hoïc vaø hoùa lyù cho saûn phaåm. Haøm löôïng acid deã bay hôi taïo thaønh bôûi naám men coá ñònh thaáp hôn, do ñoù chaát löôïng saûn phaåm ñöôïc caûi thieän. Chöông 5: Keát luaän vaø kieán nghò KEÁT LUAÄN VAØ KIEÁN NGHÒ KEÁT LUAÄN Töø caùc keát quaû treân, chuùng toâi nhaän thaáy raèng, haøm löôïng ñöôøng vaø tannin ñeàu coù aûnh höôûng lôùn ñoái vôùi ñoäng hoïc quaù trình leân men röôïu vang. Nhìn chung, khaû naêng sinh tröôûng (theå hieän qua maät ñoä teá baøo cöïc ñaïi, toác ñoä sinh tröôûng rieâng cöïc ñaïi) cuûa naám men coá ñònh thaáp hôn so vôùi naám men töï do. Chæ tröø tröôøng hôïp khi söû duïng haøm löôïng tannin ban ñaàu cao (9,8 – 17,8g/L) thì khaû naêng sinh tröôûng cuûa naám men coá ñònh môùi cao hôn do luùc naøy khaû naêng sinh tröôûng cuûa naám men töï do bò öùc cheá raát nhieàu. Tuy nhieân, so vôùi naám men töï do thì chuùng toâi thaáy raèng naám men coá ñònh laïi coù nhieàu öu ñieåm: Thôøi gian leân men ruùt ngaén raát nhieàu. Toác ñoä söû duïng ñöôøng vaø toác ñoä sinh toång hôïp coàn cao hôn; ñaëc bieät toác ñoä söû duïng ñöôøng rieâng vaø toác ñoä sinh toång hôïp coàn rieâng thì cao hôn nhieàu. pH vaø haøm löôïng acid toång thaáp hôn, do ñoù giuùp caûi thieän ñoä beàn hoùa lyù vaø sinh hoïc cho saûn phaåm. Haøm löôïng acid deã bay hôi thaáp hôn, do ñoù giuùp caûi thieän chaát löôïng saûn phaåm. Ngoaøi ra, vieäc öùng duïng naám men coá ñònh trong saûn xuaát röôïu vang coøn coù öu ñieåm khaùc laø khaû naêng taùi söû duïng raát cao. KIEÁN NGHÒ Do nhöõng haïn cheá veà thôøi gian vaø kinh phí neân nghieân cöùu cuûa chuùng toâi chöa khaûo saùt heát ñöôïc taát caû caùc vaán ñeà lieân quan ñeán ñoäng hoïc quaù trình leân men röôïu vang söû duïng naám men coá ñònh trong gel alginate. Töø ñoù chuùng toâi ñöa ra moät soá kieán nghò sau: Khaûo saùt theâm ñoäng hoïc hình thaønh caùc hôïp chaát höông. Khaûo saùt theâm aûnh höôûng cuûa caùc yeáu toá coâng ngheä quan troïng khaùc nhö: maät ñoä caáy, nhieät ñoä leân men, haøm löôïng nitô ban ñaàu ñeán ñoäng hoïc quaù trình leân men röôïu vang. Khaûo saùt khaû naêng taùi söû duïng cuûa naám men coá ñònh trong chaát mang alginate. Toái öu hoùa quaù trình leân men vaø tieán haønh saûn xuaát thöû ôû quy moâ lôùn. Khaûo saùt vieäc coá ñònh naám men treân caùc loaïi chaát mang môùi, ñaëc bieät laø caùc chaát mang traùi caây nhö taùo, leâ, moäc qua,.. Taøi lieäu tham khaûo TAØI LIEÄU THAM KHAÛO Buøi AÙi, Coâng ngheä leân men öùng duïng trong coâng ngheä thöïc phaåm, Nhaø xuaát baûn Ñaïi hoïc Quoác gia TpHCM, 2003, 235p. Phaïm Troïng Khoa, Nghieân cöùu quaù trình leân men coàn baèng naám men coá ñònh trong gel alginate, Luaän aùn cao hoïc, 2002. Leâ Vaên Vieät Maãn, Laïi Mai Höông, Thí nghieäm vi sinh vaät hoïc thöïc phaåm, Nhaø xuaát baûn Ñaïi hoïc Quoác gia TpHCM, 2006, 152p. Offiical methods of analysis of AOAC international, 1996. 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