Supplementary Material MEP-based biogeochemistry modeling. Vallino, J.J.

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1 Supplementary Materal, Vallno, J.J. S Supplementary Materal MEP-based bogeochemstry modelng. Vallno, J.J. Ths supplementary materal provdes addton detals on modelng as well as prelmnary data on the mesocosms experments beng conducted to test the MEP hypothess and modelng approach. Metabolc Hal Reactons The explct orm o the hal reactons dsplayed n Fg. n the man text s gven n Table S. Table S. Hal reactons used or the methanotroph metabolc network, where S s bologcal structure wth a C:N rato o ρ S, ac and an are the racton o assmlated C and N, and dc and dn are detrtal C and N, respectvely. Reacton Structure Rate CH + H O CH O + e - + H + S r CH O + H O CO + e - + H + S r O + e - + H + H O S 3 r 3 NO e - + H + NH + + 3H O S r CH O + NH + /ρ S +.7e - S S 5 r 5 S ac CH O + (- ac )dc + ( an NH + + (- an )dn) /ρ S +.7e - S 6 r 6 dc CH O S 7 r 7 dn NH + S 7 * r 8 * Note, S 7 catalyzes both reactons 7 and 8. State Equatons For the methanotrophc example, methane n ar s sparged nto a mcrocosm o volume V L at low rate F g wth gas partal pressures o p CH (.9 atm), po (. atm) and p CO ( atm) (Fg. S). The mcrocosm s ntally augmented wth nutrents (7 µm NO 3 -, µm NH +, and

2 Supplementary Materal, Vallno, J.J. S µm CH O) and.5 µm S T, but s otherwse closed except or gas transport. The explct orm o the state equaton (Eq. ) s gven by, dcch = r + kla( pch / κ CH ( T ) cch ( VL (S) dc O = r r r5 + r6 r7 ( ac ) (S) dc NO 3( = r (S3) dc = r + ( r6 r5 r8 ( an ρ S ) (S) dco = r3 + kla( po / κ O( T ) co( VL (S5) dcco( = r + kla( pco / κ CO( T ) cco( VL (S6) dcdc = ( ac ) r6 r7 (S7) dcdn = ( an ) r6 / ρ S r8 (S8) ds = σ r5 S r6 / ST, or =,,7 (S9) dζ ATP = / S T 8 = η r (S) dζ NADH 8 = / S ε r (S) T =

3 Supplementary Materal, Vallno, J.J. S3 ( pch pch + klart ( cch ( pch ( / CH ( T ) dpch = Fg κ (S) ( po po + klart ( co( po( / O( T ) dpo = Fg κ (S3) ( pco pco + klart ( cco pco / CO ( T ) dpco = Fg κ (S) where κ (T ) s the Henry's law constant or gas, k L s the lqud-sde mass transer coecent, a s the ar-gas nterace area, and V G s the reactor gas volume. Expermental Data Four dentcal 8 L mcrocosms (MCs) consstng o a dened mneral salts medum (7 µm NO - 3, 7 µm PO 3-, plus salts and trace elements, ph 6.8) were each noculated wth L o whole water collected rom a cedar swamp located n Woods Hole, MA on 7 Oct 5, and renoculated on day 3. The mcrocosms (Fg. S) have been operatng contnuously or 7 months and are sparged at ml mn - (STP) wth a gas mxture o.9% methane n ar. The mcrocosms are gently mxed wth a cell culture mpeller ( RMP) and mantaned n the dark at C. Other than or occasonal samplng or chemcal analyss (Fg. S) and assocated medum replacement, the reactors are closed except or gas transport, whch s contnuously montored or CH, CO and O composton (Fg. S3).

4 Supplementary Materal, Vallno, J.J. S Fg. S. Methanotrophc mcrocosm (MC o ) sparged wth.9% methane n ar ater 7 months o contnuous operaton. The mcrocosms are perodcally sampled or dssolved and partculate consttuents. Note the accumulaton o partculate matter at bottom o reactor.

5 Supplementary Materal, Vallno, J.J. S5 (a) (µm) NO 3 - NH + (µm) 8 MC MC MC3 MC 6 8 Tme (d) (b) TDN (µm) DOC (mm) MC MC MC3 MC 6 8 Tme (d) Fg. S. Prelmnary methanotrophc mcrocosm data or our replcates llustratng changes n (a) ntrate and ammonum concentratons and (b) dssolved organc carbon (DOC) and total dssolved ntrogen (TDN). Note, DOC s plotted as mm. Dssolved organc ntrogen (DON) s obtaned by subtractng NO 3 - and NH + concentratons rom TDN concentraton. Medum was added on days 63 ( L), 3 ( L), and 5 (.65 L) to replace volume o samples removed or analyses.

6 Supplementary Materal, Vallno, J.J. S6..8 Carbon Doxde (%) Feed Gas MC MC MC 3 MC 3 Oxygen (%) Tme (d) Methane (%) Tme (d) Tme (d) Fg S3. Gas concentratons (%) o CO, O and CH enterng (purple lne) and extng each o the our replcate mcrocosms. All mcrocosms were renoculated wth whole water samples on day 3 because o lkely extncton events assocated wth ntal noculum. Followng renoculaton, mcrocosm dynamcs stablzed, but dd respond to.65 L o medum added on day 5.

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