: .

: 11/06/2015 :

1. 1.1

, . SO2 , 5% .

:

SO2 + H2O H2SO3 H+ + HSO3-HSO3- H+ + SO32- , pH .

. . pH , pKa1 pKa2, 1,8 6,9 . pH 0 2 2 7 pH 7 9. HSO3-. 1.1 SO2 pH. pH.

.

pH SO2 (mg/L)% SO2

2.84.649.28

3.03.036.06

3.21.963.91

3.41.252.51

3.60.801.60

3.80.511.01

4.00,320,64

4.20,200,41

1.1 pH SO2, 50 mg/L .

, , , , . , , - (Rankine and Pocock 1969), . , 50-200 mg/L .

, , .

. , ( 1974). , SO2 . , SO2 SO2 Lineweaver-Burk. , SO2 ( 50C ) pH ( pH SO2. SO2 , ( Stratford & Rose 1986). SO2 . . , . , pH . . -. pH . pH . , pH 5.5-6.5, ( Imai and Ohno 1995), HSO3- SO32-. SO2, SO2 . , , SO2 , (Stratford & Rose 1986) ( 1974; Pilkington and Rose 1986). , , , HSO3- - , , , HSO3- (Park and Bakalinsky 2004). , Sul1p Sul2p . , ATP, pH. SO2, SO2 0.003. , :

.

.

.

.

, pH, .

1.2 . , , pH HSO3- . , . , S. cerevisiae. , , , , . ( Baldwin 1951).

, , . , , , . (Ventre 1934). , , . (Schimz 1980). , . , , . SO2, (Anacleto and van Uden 1982):

a) , SO2, .b)

c) ATP , ATP. ( Freese et al. 1973; Schimz and Holzer 1979; Schimz 1980; Prakash et al. 1986). , , . , . - , . .

1.1.1 , (CSH) (CSSC) ( ). .

, , .

, , (EC 1), , . , NAD + / NADP+ ( EC 1.1.1), (EC 1.1.2), (EC 1.1.3), (EC 1.1.5) (EC 1.1.99). , Zn, , , NAD+ . To , NADH2. , NADH2 NAD+. , . , . NAD(P)+. SO2 3- 2 ATP , ATP. Maier (1986), , , ATP 70%. , , ATP NAD+ , . (Stratford and Rose, 1986, Divol et al., 2012). , , (, -), , , .

, :

CH3CHO + SO2 + H2O CH3CHO-HSO3- + + , NADH2 , 3P- . , NADH2, , 3P- (DHAP) . . , . 7-10% . , NAD ,FAD, FMN, (Carnack et al. 1950).

ATP . ATP. (12) . , , ALD6, . , MET16, PAPS, ( Aranda et al. 2006). , , / C/G ( Mukai et al. 1970; Pagano and Zeiger 1987; Pagano et al. 1990; Meng and Zhang 1992).

. . , . . pH.

1.3 . , (Richmond, 1973). , , . -2 +6. , , +6. ( Huxtable, 1986; Kelly, 1988). , , S2- , .

. S. cerevisiae, 0,2-0,9% (Maw, 1963,1965). , 0,14% (Phaff, 1971). , (Hartnell and Spedding, 1979). H , - (Karlson, 1988). S. cerevisiae, : 0,13-0,27 % , 0,15-0,28% 0,05-0,10% (Maw, 1965). , 20% , 40% 90% . , , -CoA, , . Saccharomyces spp., 29-90 g/g . S. cerevisiae 16,9 g/g . S. cerevisiae Candida utilis, S-, 0,5 2,5 moles/g . Saccharomyces spp., 0,5-4,5 g/g . (Maw, 1965)

, , . S35 , 1,3% 3.0% . Maw (1965), , , 60% , 5% . , , , , . 1.4 , . . , , , . (Maw, 1960; Hysert and Morisson, 1976; Breton and Surdin-Kerjan, 1977). , ( Cartwright et al., 1989). O McCready Din (1974), S. cerevisiae SulP . Kotyk (1959), Maw(1963) Roomans (1979), , Michaellis-Menden. m Vmax, pH, . , Breton Surdin-Kerjan (1977), , S. cerevisiae, , Lineweaver-Burk, . , 5- (APS), . , . Maw (1963), , , , , . , , . H . , L- , L-, L- DL-, . , , . , , . . , 2,4- , . ( , 1963; McCready & Din, 1974; Cooper,1982).

Saccharomyces spp., . , . . S. cerevisiae (Catwright et al., 1989). , . Saccharomyces, , Candida utilis, S. cerevisiae ( Suomalainen and Oura, 1971). , Saccharomyces. , , , . , 2-, , DL- . , . , S--L-, , (Maw, 1965). 1.5

, . , , ( aw, 1965). 10 mg/L , S. cerevisiae. , . Candida utilis, . Saccharomyces, , . , . , 100g/L , 1 g/L. , 50-500 g/L (Maw 1965). C18 , DNA-RNA, . . , . 1.6 .

, , . .

, . . S. Cerevisiae .1.7 , S2- (De Robichon-Szulmajster and Surdin-Kerjan, 1971, Jones and Fink, 1982). 1.7.1. Gibbs, S2- 800 kJ/mol (Huxtable, 1986), .

[1], APS. , Sul1p/Sul2p, . o 5- (ATP) 5- (APS) [2]. ATP. , P. , Km , , PS. (Slaughter, 1989). APS 3- 5- (PAPS) [3] APS (Robbins and Lipmann, 1958). H PAPS , (Baumgarten, 1981. Bidan and Collon 1988). PAPS 3,5- (PAP), PAPS NADPH + H+ [4] (Peck and Lissolo, 1988). , (Asahi et al., 1961; Wilson et al., 1961; Wilson and Bierer, 1976). in vitro , , PAPS, . , , . C, , . ( Wilson et al, 1961; Maw 1965; Lawrence and Cole, 1968). , (SO32-), (Hilz et al., 1959; Hilz and Kittler, 1960), C. , C, NADPH. , , , C NADPH ( 1.7.2). , PAPS, . (Lawrence and Cole, 1968; Baumgarten, 1981; Bidan and Collon, 1988). C, , +6 +4. +6 +5 , , +4 . PAPS, - PAPS. ( Baumgarten, 1981; Peck and Lissolo, 1988).

1.7.1 [ 1. ( ); 2. ATP ( EC* 2.7.7.4); 3. APS (EC 2.7.1.25); 4. PAPS 5. ;

6. (E.C. 1.8.1.2); 7. (E.C. 2.3.1.30); 8. (E.C. 4.2.99.8); 8b. - (E.C. 4.2.99.10); 9. - (E.C. 4.2.1.22); 10. (E.C. 4.4.1.1); 11. (E.C. 2.3.1.31); 12. (E.C. 4.2.99.10); 13. - (E.C. 4.2.99.9); 14. EC (4.4.1.8); 15. EC (2.1.2.1); 16. 5,10--(EC1.1.99.15);

17. ( EC 2.1.1.14); 18. SAM ( ) ( EC 2.5.1.6); 19. SAM; 20. ( EC 3.3.1.1)].

1.7.2 3- 5- (PAPS). fraction C, .

PAPS in vitro, C, - PAPS ( Wilson et al., 1961; De Robichon-Szulmajster and Surdin-Kerjan, 1982; Bidan and Collon, 1988). [5] ( Dott and Truper, 1978 c; Stratoford and Rose, 1986). , PAPS, , , -, . (Tori and Bandurski, 1967).

S2- [6]. Yoshimoto Sato, (1968 1970), , , , NADPH . , , FMN, FAD, 587 nm, , . Prabhakararao Nicholas (1969, 1970), , . , , , sirohaem. -, , . To NADPH sirohaem (Lehninger, 1977; Karison, 1988). sirohaem [4Fe-4S].(Peck and Lissolo, 1988). Cooper (1983) Slaughter (1989), , , NADPH, , . . S. cerevisiae, . - (Umbarger, 1978).

1.7.4 PAPS, . 1. - ; 2. ; 3. (E.C. 1.6.4.5); 4. ( EC 1.8.1.2) , ( 1.7.4), PAPS, - PAPS ( S-SO3-), , NADPH (-S-S-). , , - - (Jones and Fink 1982; 1.7.1). , , . ADP-, , APS , ADP . . 3- , PAPS APS in vitro .( Bidan and Collon, 2009). 1.8 ( 1.7.1).

(Jones and Fink, 1982; Ono et al., 1988). , [7] -. ( 8a + b), , (Tsang and Schiff, 1976). -. , (Jones and Fink, 1982), , . [12] (Yamagata and Takeshima, 1976; Yamagata, 1989). , - [9]. , - [10]. Schlossmann Lynen (1975), , . Dott Truper (1978), , . , S. cerevisiae (Yamagata 1989). , - [11] (Nalki and Yamagata, 1973). - . - [12]. , . , - - [13]. -, , , . [14]. (Yamagata, 1989). , [17]. , 5-- (THF) (Jonew & Fink, 1982). , [15] 5,10- [16].

(Ono et al., 1988). trans- , SAM (S-) SAM [18]. H SAM S SAM [19]. S-, [20] . , - [9] (Delavier-Klutchka and Flavin, 1965). To 2--, - [10] (De Robichon-Szumajster and Surdin-Kerjan, 1971; Jones and Fink, 1982).

, . , , , SAM, . ( Cherest et al., 1973; Jones and Fink 1982). H , -tRNA , SAM . ( Masselot and de Robichon-Szulmajster, 1972; Surdin-Kejal et al. 1972). . - . , -, - -. SAM . H 5,10--THF. SAM (Botsford and Parks, 1969; Jones and Fink, 1982).

ATP-, (De Vito and Dreyfuss, 1964). , , . (Baumgarten, 1981).

S. cerevisiae, , , , ( ), (Cherest et al., 1969). 1.9

. Nwoguh Berry (1990), , Saccharomycopsis fibuligera. . . SAM (Slaughter, 1989).

. , , . , , . , , , , Na2SO3, Na2S, Na2SO4, S- (Umezawa and Kishi, 1989). , Rhodotorula glutinis, , . (Maw, 1985; Umezawa and Kishi, 1989).

.. - . (Slaughter, 1989).

1.10 SO2 SO2 , . , Thorton (1982), SO2 , , SO2 . , SO2 SO2 (Thomas 1985). To 1994, , , SO2, SO2 (Xu et al. 1994). H SO2, ( Casalone et al. 1992). SO2, , (Steels et al. 2000). H , ATP ( Warth 1977 ). SO2 , 80, SO2, , . , Zygosaccharomyces baili SO2 (Warth 1977; Thomas and Davenport 1985; Warth 1985; Pilkington and Rose 1988; Divol et al. 2006), Schizosaccharomyces pombe (Stratford et al. 1987). Kloeckera apiculata Hansenula anomala, , SO2 (Warth 1985). , S. cerevisiae, , SO2 (Divol et al. 2006; Nardi et al. 2010). , SO2, 50 mg/L, non Saccharomyces , Candida spp., SO2, 20 mg/L, non Saccharomyces (Henick-Kling et al. 1998).1.10.1 SO2.

O S. cerevisiae, SO2 (Park & Bakalinsky 2000). , , , SO2 : ) 1.7 S2- ( Yoshimoto & Sato1968; Kobayashi and Yshimoto 1982), ) SO2 (Heimberg et al. 1953; Beck-Speier et al. 1985; Kappler and Dahl 2001; Friedrich et al. 2005; Feng et al. 2007; Hellborg and Piskur 2009), ) ( Stratford et al. 1987) ) (Mannervik et al. 1974; Katgedal et al. 1986) ) SO2 Ssu1p (Park and Bakalinsky 2000). 1.10.1.1 SO2 SO2 , (Charles & Suzuki 1966; Suzuki & Silver 1966; Silver and Lundgren 1968). S. cerevisiae (Beck-Speier et al. 1985; Xu et al. 1994) Rhodotorula (Kurek 1985), , , . ( Benoit Divol; Maret du Toit; Edward Duckitt 2012).

1.10.1.2 , , , ( 100 ppm), , , . ( Bartwsky & Pretorius 2009).

, 1.2. SO2, 1.10.1. S. cerevisiae SO2, SO2, . (Stratford et al. 1987; Casalone et al. 1992) , ( Casalone et al. 1992; Divol et al. 2006). , . , SO2 . , , SO2, : ) , ) SO2 ( Frivik and Ebeler 2003). , , .

1.10.1.3 H SO2 . (Katgedal et al. 1986; Casalone et al. 1989), , SO2, ( Mannervik et al. 1974; Katgedal et al. 1986) :GSSG + HSO_3 GSSO_3 + GSH

, GSSG , GSH GSSO-3 S- . , S. cerevisiae GLR1. , SO2, GSH (Casalone et al. 1992) GSH SO2 (Xu et al. 1994). , .1.11 , SO2, . Park Bakalinsky (2000), SO2 . , , SO2, .

, , . FZF1 S. cerevisiae (Casalone et al. 1994). (Avram & Bakalinsky 1996). , FZF1 SO2, GRR1. GRR1 CAT80, COT2, SSU2, SDC1. , (Bailey & Woodword 1984; Flick & Johnston 1991; Erickson & Johnston 1994), (Conklin et al. 1993), (Kim et al. 1994) tutnover G1 (Barral et al. 1995). GRR1 . SO2 , . GRR1 SO2 , . , . , . Bakalinsky & Avram (1996) , , , SO2 . FZF1, SSU1, SSU1 [-445; -378] (Avram & al. 1999). Fzflp 11 - , Fzflp, SSU1. SSU1, XVI. GRR1, SO2 ( Avram & Bakalinsky 1997; Park & Bakalinsky 2004). , Ssu1p, , SO2 . , . SO2, SSU1, SSU1-R, 97% SSU1. (Aa & al. 2006). ( ) SSU1 (. XVI) ( Nardi et al. 2010), . (. VIII), (Aa et al. 2006; Liti et al. 2009; Nardi et al. 2010). S. cerevisiae, , SSU1 SSU1-R .

, , VIII (Goto-Yamatoto et al. 1998; Perez-Ortin et al. 2002). , SSU1-R, ECM34 ( 1.11.1). SSU1-R, SSU1 (Goto-Yamatoto et al. 1998). To SSU1-R, SSU1 Fzflp. , Fzflp [-442; -420], (9bp) SSU1 (76 bp) SSU1-R (Yuasa et al. 2004). , Yuasa & al., 2005, SSU1, SO2. , , 76-bp SSU1-R, SSU1-R. SSU1-R . , . , 76-bp SSU1 SSU1-R, . , FZF1, SSU1, SSU1- R. , . , SSU1, FZF1 ( Sarver and Derisi 2005). , FZF1, , CS2, , CS1, ( 1.11.1) , SSU1 SSU1-R (Yuasa et al. 2005; Aranda et al. 2006; Park & Hwang 2008). , , PCR, SSU1, (Nardi et al. 2010). FZF1, ( SSU1, SSU1-R. , . SSU1, (Divol et al. 2006; Nardi et al. 2010). , , SSU1, SSU1-R (Nardi et al. 2010). , SSU1 SO2, , SSU1-R. - Ssu1p , Nardi & al., 2010, SSU1. , SO2.

, SSU1-R ( Perez-Ortin et al. 2002).

1.12 , . , , , .

SO2 , 21 , . . , PDC1. , . , TDH3, 3- , , ADH1 . (Maier et al. 1986). , PDC1, ADH1, , , SO2. , SO2 (. FZF1, SSU1), (MET1, MET5, MET8, MET10) (GLR1), . 1.11 O S. cerevisiae SO2, 10-30 mg/L. , - 100 mg/L. .

, . , .

, .

, S. cerevisiae . , .1.12 , 1.7, S2- . . , 50 80 g/L. . . S. cerevisiae. 1 mg/L . . pH .

. , .

. , , , .

. .

2.

2.1

20 ppm . , : , pH, DNS.

2.1.1 1 ml 4 ml 0,1 (). :gr/L = * 0,75 * 10

2.1.2 DNS

0, 0.5, 1, 1.5, 2 540 nm .

C

0,0070

0,2310,5

0,4431

0,7451,5

0,7842

2.1.2.1 540 nm .

2.1.2.1

, 2.1.2.1, . 1 ml , . 0,3-0,8 1/100 . , 1 ml 1 ml . 1 ml DNS . 5 . 2 10 ml . 540 nm. 2.2 (h)0120168288336

(g/L)1,723,07533,564,1

PH43,233,113,09

(g/L)200133,873,42,820,9298

(g)335,93324,65314,65282,36260,51

2.2.1 20 ppm . 2.2.1 20 ppm . 3 2.2.1 . T . 120-168 h ( 1,26 g/h). , , , , , .

.

, . , , .

, , 2.2.1, 288-336 h ( 0,45 g/h ). , , . , , .

pH , , .

, , , , , - ., . , 2.2.1

pH, 168h . ,