Data_Run1

Selected Cell

Cell:

Value:

Run1_CF8_data PHA_assay Carbon_assay Nitrogen_assay PumpCalibration

Exp	Run 1 [CF8]									AntiFoam addition	Dilution rate	Time [h]	OD [540 nm]	DO%	N [mg/l]			Graph												Volumetric productivity		Biomass to Substrate ratio [Cmol/Cmol]
												-1		100	283.5820154998324			Time [h]	100*CDW [g/L]	AA [mM]	BDO [Mm]	TA [Mm]	EG [mM]	N [mg/L]	Dilution rate	Time [h]	CDW [g/L]	PHA [% CDW]	PHA [mg/l]	CDW [mg/l/h]	PHA [mg/l/h]	Substrate [Cmol]	Biomass C [g/l]	Biomass [Cmol]	Ratio [%]
												0	0.137	93	259.83973405812833		Theoritical concentrations	-1		9.06	13.19	3.4	13.6	283.5820154998324	0.1	24	0.44799999999999995	0.34	1.5231999999999999	44.8	0.15232	0.16318374823078846	0.234712576	0.01954146832070602	11.975131428571428
Bacteria and Monomers	P. putida KT2440 ΔΔP14e_dcaAKIJP	Adipate [AA]										2	0.2	91	246.6001628133177			0		8.658846265362037	13.200525954572909		13.45369245992038	259.83973405812833	0.1	97	0.4666666666666666	0.65	3.033333333333333	46.666666666666664	0.30333333333333334	0.16318374823078846	0.24449226666666665	0.020355696167402102	12.474095238095236
	P. putida CA3 - ALE	1,4-butanediol [BDO]										4	0.235	88	236.9863071413141			2		9.982757037593597	14.964702905452185		14.307071251507944	246.6001628133177	0.1	122	0.4606060606060606	0.67	3.086060606060606	46.06060606060606	0.3086060606060606	0.16318374823078846	0.24131704242424243	0.020091336476916365	12.31209400123686
	P. umsongensis GO16	Ethylene glycol [EG][										6	0.537	79	215.81704985579086			4		10.43707564223735	14.78953533148543		14.013017156079098	236.9863071413141	0.075	200	0.49375	2.59	12.788125	37.03125	0.959109375	0.16318374823078846	0.25868155000000004	0.021537053534260265	13.198038265306122
		Terephthalate [TA]									0.1/h from T 7:00	24	1.7730000000000001	68	23.492713579167955			6		9.453997960273117	12.676027346022984		11.866852815878921	215.81704985579086	0.056	314	0.475	2.42	11.495	26.599999999999998	0.64372	0.16318374823078846	0.2488582	0.02071919074182	12.696846938775511
										Precautionary AF T32		97	1.6300000000000001	20	1.4236795767942474			24	44.8	8.75249064207949	5.205825664836	0	3.077282487276659	23.492713579167955	0.1	367	0.4702127659574468	1	4.702127659574468	47.02127659574468	0.4702127659574468	0.16318374823078846	0.2463501106382979	0.020510374709707593	12.568883195831523
Monomer plates	2024-02-02T0:0:0+0000											122	1.6300000000000001	26	0.9476640429660715			97	46.666666666666664	5.2582490616010835	5.671081585715886	0	3.7216819447755567	1.4236795767942474	0.075	462	0.4954545454545455	1.32	6.540000000000001	37.159090909090914	0.49050000000000005	0.16318374823078846	0.2595745818181818	0.021611404697209378	13.24360111317254
Inoculum prep											0.075/h from T 124	200	1.927	52	0.20288089212508978			122	46.060606060606055	4.827479135667906	5.353028681995796	0	3.762265874974386	0.9476640429660715	0.056	582	0.47600000000000003	1.44	6.854400000000001	26.656000000000002	0.3838464000000001	0.16318374823078846	0.24938211200000004	0.02076281009075015	12.723577142857145
2024-12-02T0:0:0+0000	1899-12-31T17:0:0+0000	12.5 ml MSM fN + monomer [TA + EG for GO16]	3 bugs as Monocultures								0.056/h from T 201	314	1.78	60	0.12756048525415675			200	49.375	1.7208204209043907	8.81998105238108	0	7.087835903781801	0.20288089212508978	0.056	677	0.6673469387755102	14.48	96.63183673469389	37.37142857142857	5.411382857142858	0.2031804179502123	0.3496310693877551	0.029109238979914673	14.326793533345153
13/2/2024	1899-12-31T16:30:0+0000	50 ml MSM Fn Monocultures									0.1/h from T 315	367	1.588	50	0.13691705872699134			314	47.5	0.9604848536810233	7.676647386016818	0	5.822637859783717	0.12756048525415675	0.056	792	0.696	16.81	116.99759999999999	38.976	6.551865599999999	0.2031804179502123	0.364642752	0.03035906685538257	14.941925585969514
14/2/2024	1899-12-31T18:0:0+0000	50 ml MSM Fn Monocultures									0.075/h from T 367	462	1.723	47	0.14112751766116555			367	47.02127659574468	1.127266270674248	10.430378043655836	0.014964658693203627	8.762540952693808	0.13691705872699134
	OD at T23		Vol [uL] for 0.005 OD - 50 ml								0.056/h from T 462	582	1.8760000000000001	59	0.011305080332055777			462	49.54545454545455	1.6828760554543865	9.218363024128342	0	7.094214517565506	0.14112751766116555
13/2/2024	aleCA3	2.09	119.61722488038278								0.056/h + 5mM Octanoate from T 583 [C:N of 30:1 maintained by increasing N source accordingly]	677	2.8000000000000003	46	0.05177225929496733			582	47.6	2.42105327186153	9.000319810102082	0	6.540152177778345	0.011305080332055777
	GO16	1.88	132.97872340425533									792	2.926	53	0.16943113890981082			677	66.73469387755102	2.5429097187685286	11.561662349816022	0	9.68823727805414	0.05177225929496733
	KTJP	1.45	172.41379310344828															792	69.6	2.1462897217403545	12.326545694437263	0	11.269336169690263	0.16943113890981082
	OD at T25		Vol [uL] for 0.05 OD - 50 ml
14/2/2024	aleCA3	3.23	773.9938080495356
	GO16	1.77	1412.4293785310733
	KTJP	2.82	886.5248226950355
																														Graph
15/2/2024	OD at T16		Vol [mL] for 0.0333 OD x 1 L																											MEAN
	aleCA3	3.82	8.726003490401396																											Biomass and PHA result				Volumetric productivity
	GO16	2.17	15.360983102918588																									PHA [mg/L]		Dilution rate	CDW [g/L]	PHA [% CDW]		Dilution rate	CDW [mg/l/h]	PHA [mg/l/h]
	KTJP	3.5	9.523809523809524																									3.06		d = 0.1	0.4636363636363636	0.66		d = 0.1	46.36363636363636	0.305969696969697
																												9.669474431818182		d = 0.075	0.49460227272727275	1.955		d = 0.075	37.09517045454545	0.7248046875
																												9.177150000000001		d = 0.056	0.47550000000000003	1.93		d = 0.056	26.628	0.5137832
																												106.64781428571429		d = 0.056 [+ 5 mM oct]	0.6816734693877551	15.645		d = 0.056 [+ oct]	38.17371428571428	5.9816242285714285


																														Graph
																														SD
																														Biomass and PHA result				Volumetric productivity
																														Dilution rate	CDW [g/L]	PHA [% CDW]		Dilution rate	CDW [mg/l/h]	PHA [mg/l/h]
																												6.060606060606049E-4		d = 0.1	0.004285495643554822	0.014142135623730963		d = 0.1	0.4285495643554805	0.0037283812098926976
																												0.010823863636363656		d = 0.075	0.0012052956497497999	0.8980256121069148		d = 0.075	0.09039717373123797	0.33135686679008963
																												0.0049000000000002765		d = 0.056	7.071067811865874E-4	0.6929646455628166		d = 0.056	0.03959797974644984	0.18375838481136042
																												0.33380816326530516		d = 0.056 [+ 5 mM oct]	0.020260773893590017	1.6475588001646544		d = 0.056 [+ oct]	1.1346033380410459	0.8064430813005183



Time [h]	CDW [mg]	Volume [mL]	CDW [g/L]	PHA sample [mg]	PHA %	PHA [mg/l]
24	11.2	25	0.44799999999999995	11.2	0.34	0.0015232
97	11.2	24	0.4666666666666666	11.2	0.65	3.0333333333333328
122	15.2	33	0.4606060606060606	10.8	0.67	3.086060606060606
200	15.8	32	0.49375	9.7	2.59	12.788124999999999
314	19	40	0.475	10.3	2.42	11.495
367	22.1	47	0.4702127659574468	7.8	1	4.702127659574469
462	21.8	44	0.4954545454545455	9.7	1.32	6.540000000000001
582	23.8	50	0.47600000000000003	10.7	1.44	6.854400000000001
677	32.7	49	0.6673469387755102	13.4	14.48	96.63183673469389
792	34.8	50	0.696	13.9	16.81	116.99759999999998










PHA - Monomer Composition
%
	C4 (%)	C5 (%)	C6 (%)	C7 (%)	C8 (%)	C9 (%)	C10 (%)	C12 (%)	Total PHA [as % CDW]	0
24	0	0	0	0	0	0	100	0	0.34	100
97	0	0	0	0	0	0	100	0	0.65	100
122	0	0	0	0	0	0	100	0	0.67	100
200	0	0	0	0	12.89	0	87.11	0	2.59	100
314	0	0	0	0	13.83	0	86.17	0	2.42	100
367	0	0	0	0	0	0	100	0	1	100
462	0	0	0	0	0	0	100	0	1.32	100
582	0	0	0	0	0	0	100	0	1.44	100
677	0	0	10.63	0	83.32	0	6.05	0	14.48	99.99999999999999
792	1.3	0	10.68	0	83.02	0	5	0	16.81	100

MEAN													SD
Dilution rates	Time [h]	C4 (%)	C5 (%)	C6 (%)	C7 (%)	C8 (%)	C9 (%)	C10 (%)	C12 (%)	Total PHA [as % CDW]	Cross check		Dilution rates	Time [h]	C4 (%)	C6 (%)	C8 (%)	C10 (%)	C12 (%)	Total PHA [as % CDW]
0.1	122 and 367	0	0	0	0	0	0	100	0	0.835	100		0.1	122 and 367	0	0	0	0	0	0.2333452377915609
0.075	200 and 462	0	0	0	0	6.445	0	93.555	0	1.955	100		0.075	200 and 462	0	0	9.114606409494598	9.114606409494598	0	0.8980256121069148
0.056	314 and 582	0	0	0	0	6.915	0	93.08500000000001	0	1.93	100.00000000000001		0.056	314 and 582	0	0	9.779286783809953	9.779286783809951	0	0.6929646455628166
0.056 + Oct	677 and 792	0.65	0	10.655000000000001	0	83.16999999999999	0	5.525	0	15.645	100		0.056 + Oct	677 and 792	0.9192388155425119	0.035355339059326626	0.21213203435596226	0.7424621202458748	0	1.6475588001646544


Without C5, C7, C9 and C12
Time [h]	Dilution rates	C4 (%)	C6 (%)	C8 (%)	C10 (%)	C12 (%)	Total PHA [as % CDW]
122 and 367	d = 0.1	0	0	0	100	0	0.835
200 and 462	d = 0.075	0	0	6.445	93.555	0	1.955
314 and 582	d = 0.056	0	0	6.915	93.08500000000001	0	1.93
677 and 792	d = 0.056 + Oct	0.65	10.655000000000001	83.16999999999999	5.525	0	15.645

Experiment	Run_1 [CF8] PHA assay


	Retention times [min]
	Internal standard	8.62	8.63

	C4	5.17	5.18
	C6	9.07	9.07
	C8	15.58	15.59
	C10	22.62	22.52
	C12	30.4



				GC - peak area data										Mass of PHA monomer										PHA monomer distribution
	Sample name	Sample amount [mg]	Methanolysis conversion efficiency, X	Internal standard peak	C4	C5	C6	C7	C8	C9	C10	C11	C12	C4	C5	C6	C7	C8	C9	C10	C11	C12	Total PHA [mg]	C4	C5	C6	C7	C8	C9	C10	C11	C12	Total %PHA
Run 1 [CF8] samples	24	11.2	0.85	209.8	0	0	0	0	0	0	36.4	0	0	0	0	0	0	0	0	0.031985064071045094	0	0	0.031985064071045094	0	0	0	0	0	0	100	0	0	0.3359775637714821
	97	11.2	0.85	203.9	0	0	0	0	0	0	68.4	0	0	0	0	0	0	0	0	0.06184295046591466	0	0	0.06184295046591466	0	0	0	0	0	0	100	0	0	0.6496108242217927
	122	10.8	0.85	207.8	0	0	0	0	0	0	69.5	0	0	0	0	0	0	0	0	0.061658163720176275	0	0	0.061658163720176275	0	0	0	0	0	0	100	0	0	0.6716575568646652
	200	9.7	0.85	228.3	0	0	0	0	32.2	0	230.2	0	0	0	0	0	0	0.027517163731931665	0	0.18588773386817892	0	0	0.21340489760011058	0	0	0	0	12.894344994600257	0	87.10565500539974	0	0	2.588294694967988
	314	10.3	0.85	205.3	0	0	0	0	30.9	0	203.7	0	0	0	0	0	0	0.029364542425718457	0	0.18291671987079236	0	0	0.21228126229651081	0	0	0	0	13.832847095426901	0	86.1671529045731	0	0	2.42468603422628
	367	7.8	0.85	181.8	0	0	0	0	0	0	65.3	0	0	0	0	0	0	0	0	0.06621717179086328	0	0	0.06621717179086328	0	0	0	0	0	0	100	0	0	0.9987507057445444
	462	9.7	0.85	183.4	0	0	0	0	0	0	107.9	0	0	0	0	0	0	0	0	0.10846095861791882	0	0	0.10846095861791882	0	0	0	0	0	0	100	0	0	1.3154755441833699
	582	10.7	0.85	180.9	0	0	0	0	0	0	128.8	0	0	0	0	0	0	0	0	0.13125885799074802	0	0	0.13125885799074802	0	0	0	0	0	0	100	0	0	1.4431979987987689
	677	13.4	0.85	184.5	0	0	151.4	0	1299.6	0	99.9	0	0	0	0	0.17532679497413156	0	1.3742540956097558	0	0.09982066136071888	0	0	1.6494015519446061	0	0	10.629721717396553	0	83.31834622015131	0	6.051932062452145	0	0	14.481137418302074
	792	13.9	0.85	191	19.5	0	189.5	0	1614	0	102.8	0	0	0.025873539267015706	0	0.21197987720133268	0	1.648632552879581	0	0.09922270730228712	0	0	1.9857086766502166	1.3029876724244853	0	10.675275768998064	0	83.02489545751169	0	4.996841101065765	0	0	16.80667521498279



TA	2024-08-03T0:0:0+0000		RT = 3.98 min
Agilent 2	TA concentration [mM]	Peak area				7283.828282587063	180.6314726368164
						130.13403767126698	76.6728558901827
	5	26543.459				0.9987248295186366	134.82889971548278
	2.5	15818.067				3132.835472950382	4
	1.25	9174.033				5.695129036823488E7	72715.32879395084
	0.625	4933.127
	0.3125	2524.406
	0.15625	1335.823
	0.078125	676.709
	0.0390625	364.737



BDO	26/2/2024		RT = 24.98 min
Shimadzu	BDO concentration [mM]	Peak area
	395.9	7882389				19918.07308126964	-4121.536018589279
	197.95	3933741				20.44017033975166	3303.627509379633
	98.975	1971526				0.9999936813705892	7404.361829554661
	49.4875	991373				949567.0181250148	6
	24.74375	479004				5.205960735092738E13	3.289474446177963E8
	12.371875	234258
	6.1859375	122248
	3.09296875	62013


EG	26/2/2024		RT = 19.99 min
Shimadzu	EG concentration [mM]	Peak area
	600	7520026				12541.91005458605	-7267.732765298337
	300	3755859				23.268823254188405	5699.630903546423
	150	1877018				0.9999793479218851	12774.48180369891
	75	910084				290521.66344534344	6
	37.5	452867				4.740947064605988E13	9.791243121182075E8
	18.75	231080
	9.375	120815
	4.6875	65611


AA	26/2/2024		RT = 20.53 min
Shimadzu	AA concentration [mM]	Peak area
	100	3448624				34524.23174812047	-16317.911696359282
	50	1708573				246.06049749457682	10045.301104842469
	25	805661				0.9996953124874963	22514.355464077726
	12.5	396469				19686.306884180427	6
	6.25	199057				9.978914190266098E12	3.041377211777079E9
	3.125	103331
	1.5625	54901
	0.78125	30715


RTs		20.5 min		25.1 min		3.98 min		19.99 min
CF8	Sample	AA		BDO		TA		EG
26/2/2024		Peak area	mM	Peak area	mM	Peak area	mM	Peak area	mM
	Theoritical -cf8 - 0]		9.06		13.19		3.4		13.6
Inoculum	CF8 - GO16		0		0	0	0		0
	CF8 - aleCA3		0	41696	2.093375512788137		0		0
	CF8 - KTJP	188230	5.452112907369693		0		0		0
	-1	212764	6.162744252370001	177992	8.936207172682895	19568.274	2.686537079129271	110838	8.837409931980057
	0	298940	8.658846265362037	262929	13.200525954572909	22571.926	3.0989098040206327	168735	13.45369245992038
	2	344647	9.982757037593597	298068	14.964702905452185	23187.522	3.183425253863765	179438	14.307071251507944
	4	360332	10.43707564223735	294579	14.78953533148543	22283.466	3.0593070017578667	175750	14.013017156079098
	6	326392	9.453997960273117	252482	12.676027346022984	10033.688	1.3775295078357148	148833	11.866852815878921
	24	302173	8.75249064207949	103690	5.205825664836	0	0	38595	3.077282487276659
	97	181537	5.2582490616010835	112957	5.671081585715886	0	0	46677	3.7216819447755567
	122	166665	4.827479135667906	106622	5.353028681995796	0	0	47186	3.762265874974386
	200	59410	1.7208204209043907	175677	8.81998105238108	0	0	88895	7.087835903781801
2024-07-03T0:0:0+0000	314	33160	0.9604848536810233	152904	7.676647386016818	0	0	73027	5.822637859783717
	367	38918	1.127266270674248	207753	10.430378043655836	109	0.014964658693203627	109899	8.762540952693808
	462	58100	1.6828760554543865	183612	9.218363024128342	0	0	88975	7.094214517565506
21/3/24	582	83585	2.42105327186153	179269	9.000319810102082	0	0	82026	6.540152177778345
	677	87792	2.5429097187685286	230286	11.561662349816022	0	0	121509	9.68823727805414
	792	74099	2.1462897217403545	245521	12.326545694437263	0	0	141339	11.269336169690263



Standard
2024-02-04T0:0:0+0000			BLANK	No colour

Concentration range	MSM N-full 2x dilutions starting at 26.1850 mg/l
					0.4275071511824503	0.005088745239504422	slope	intercept
	N-concentration in well [mg/l]	OD at 635nm			8.69762229597747E-4	0.0024848502739475684	se(slope)	se(intercept)
	26.185	Over			0.999979304527958	0.004998510647497078	r2	se(y)
	13.0925	over			241593.74149451416	5	F	df
	6.54625	2.803032997329712			6.036245890823205	1.249255434657083E-4	ss reg	ss resid
	3.273125	1.4098329574661255
	1.6365625	0.6971330077247619
	0.81828125	0.3534329862194061
	0.409140625	0.17773300715255735
	0.2045703125	0.09373300083446502
	0.10228515625	0.05413300104904174
	0.051142578125	0.04523299940395355
	0.0255712890625	0.06723299928474426
	0.01278564453125	0.07583299585151672




N=OD/0.427507
2024-02-04T0:0:0+0000
CF8
Time [h]	OD at 635 nm	N-concentration in well [mg/l]	Dilution	N [mg/L]
-1	1.2123329670028686	2.835820154998324	100	283.5820154998324
0	1.1108330518798828	2.5983973405812835	100	259.83973405812833
2	1.05423295803833	2.466001628133177	100	246.6001628133177
4	1.0131330520706177	2.369863071413141	100	236.9863071413141
6	0.9226329953269958	2.1581704985579084	100	215.81704985579086
24	0.10043299504089355	0.23492713579167954	100	23.492713579167955
97	0.6086329848365784	1.4236795767942474	1	1.4236795767942474
122	0.40513301201629637	0.9476640429660715	1	0.9476640429660715
200	0.08673300154972076	0.20288089212508978	1	0.20288089212508978
314	0.05453300036954879	0.12756048525415675	1	0.12756048525415675
367	0.058533001025199885	0.13691705872699134	1	0.13691705872699134
462	0.060333001692771906	0.14112751766116555	1	0.14112751766116555
582	0.004833000977516169	0.011305080332055777	1	0.011305080332055777
677	0.0221330032544136	0.05177225929496733	1	0.05177225929496733
792	0.0724329979019165	0.16943113890981082	1	0.16943113890981082

CF8	2024-02-15T0:0:0+0000			Wi	Wf	Weight[g/min]	Volume[ml/calibration]
		Addition1	Media-weight per ml			1.0104
		At 100% pump capacity	I			236.35	235.94000000000003	233.5114806017419
			II			235.5
			III			235.97
		At 20%pump capacity	I	14.4658	61.7031	47.2373	47.2065	46.72060570071259
			II	14.4658	61.6462	47.1804
			III	14.4658	61.6676	47.2018

			Acid-weight per ml		1.1691	1.1691
			I	6.1956	22.2732	16.0776		13.75211701308699
			Base-weight per ml		1.2056	1.2056
			I	6.2017	23.5699	17.3682		14.406270736562709