RawData_BA_paper_Figure4

Selected Cell

Cell:

Value:

Figure_4_overall C_data PHA_assay N_assay Figure_5 PHA_assay _flask_exp N_assay_flask_exp C_data_flask_exp

100

1 L Continuous - 100 days
Bacterium	P. putida CA3																CF6 - C/N and PHA relation
C substrate	BA-Na [1.96 g C/L]
Medium	MSM N-lim
C/N ratio	7 ratios tested
Dilution rate, d [/h]	5 rates tested
EXP name	CF6






		Run info + Monitoring by OD measurements					Measurements
		Time [h]	Time [Days]	OD 540 nm	Dilution rate	Feed C/N ratio	Time [h]	BA [mM]	BA [g/l]	C [mg/l]	N [mg/l]	Culture C/N ratio	CDW [g/l]	CDW [mg/l]	PHA [%CDW]	PHA [mg/l]
		0					-1
		24	1	5.63	0.2	10	0
		48	2	7.48	0.2	10	1
		79	3.2916666666666665	6.09	0.2	10	2
		99	4.125	6.78	0.2	10	3
		122	5.083333333333333	4.83	0.2	10	5
		144	6	4.05	0.2	10	8
		169	7.041666666666667	4.65	0.2	10	24	0	0	0	1.1574352000890495	0	1.3705882352941103	1370.5882352941103	0	0
		192	8	4.23	0.2	10	48	0	0	0	1.1667505739328246	0	1.6823529411764682	1682.3529411764682	0	0
		217	9.041666666666666	3.43	0.2	15	79	0	0	0	0.6707069167518033	0	1.5411764705882354	1541.1764705882354	0	0
		242	10.083333333333334	3.51	0.2	15	99	0	0	0	0.6497473256033096	0	1.5647058823529385	1564.7058823529385	0	0
		263	10.958333333333334	3.35	0.2	15	122	0	0	0	0.4657686921887524	0	1.388235294117638	1388.235294117638	0	0
		290	12.083333333333334	2.7	0.2	20	144	0	0	0	1.5626539622932643	0	1.2058823529411742	1205.882352941174	0	0
		314	13.083333333333334	2.63	0.2	20	169	0	0	0	1.6069019880511954	0	1.3235294117647038	1323.5294117647038	0	0
		335	13.958333333333334	2.6	0.2	20	192	0	0	0	1.4974463453868387	0	1.2705882352941213	1270.5882352941214	0	0
		363	15.125	2.3	0.2	25	263	15.690301413843422	2.2610508852419064	1.3191943116381937	0.23987087647720745	5.499601831669397	0.911764705882357	911.764705882357	0	0
		386	16.083333333333332	2.06	0.2	25	335	2.2280493415167566	0.32107305035927225	0.1873278237207649	0.29110543261797023	0.6435050766180753	0.6470588235294189	647.0588235294189	1.29	8.347058823529503
		416	17.333333333333332	2.04	0.2	25	434	5.273846209081361	0.7599876079596696	0.44340944995081355	0.28179005877419516	1.5735453971643751	0.6294117647058913	629.4117647058913	1.69	10.637058823529562
		434	18.083333333333332	2.55	0.2	25	505	9.359924591772032	1.3488119332973088	0.7869548807987817	0.2491862503209825	3.1580991318144047	0.5352941176470521	535.2941176470522	2.13	11.401764705882211
		462	19.25	1.72	0.2	30	578	7.162036715871874	1.0320853009407165	0.6021629442367545	0.16534788572700707	3.6417940367918495	0.711764705882353	711.7647058823529	2.37	16.868823529411767
		482	20.083333333333332	1.91	0.2	30	650	4.831569832327443	0.6962533706875463	0.40622415535405415	0.11877101650813184	3.420229676372614	0.8000000000000033	800.0000000000033	2.02	16.160000000000068
		505	21.041666666666668	1.906	0.2	30	720	0	0	0	-0.05589224306265028	0	1.047058823529414	1047.058823529414	1.94	20.31294117647063
		529	22.041666666666668	2.262	0.2	25	793	0	0	0	0.051234556140762756	0	1.452941176470585	1452.941176470585	0.54	7.84588235294116
		553	23.041666666666668	2.17	0.2	25	864	8.289941046246566	1.1946219544693615	0.6969938169814677	0.07685183421114414	9.06931921841363	0.43428571428571083	434.2857142857108	1.02	4.42971428571425
		578	24.083333333333332	2.37	0.2	25	939	9.829045981250175	1.4164146711280565	0.8263972249669668	0.1280863903519069	6.451873791559805	0.40000000000000036	400.00000000000034	1.15	4.600000000000004
		606	25.25	2.65	0.2	20	1009	0.8886435525113154	0.12805797913464312	0.07471453152025359	0.02794612153132514	2.673520596999665	0.4028571428571429	402.8571428571429	2.21	8.903142857142857
		628	26.166666666666668	2.75	0.2	20	1084	0	0	0	0.17699210303172588	0	0.5199999999999998	519.9999999999998	1.78	9.255999999999997
		650	27.083333333333332	2.73	0.2	20	1227	7.958924057488429	1.1469207513043702	0.6691628839032757	0.07685183421114414	8.707181692824733	0.5514285714285742	551.4285714285742	1.05	5.790000000000029
		677	28.208333333333332	3.64	0.2	15	1396	2.867752029121499	0.4132574061565536	0.2411121408201981	-0.051234556140762756	-4.706045274555755	0.46285714285714263	462.85714285714266	4.84	22.402285714285703
		700	29.166666666666668	3.76	0.2	15	1563	2.042449684237173	0.29432721174699783	0.17172315140330366	0.13740176419568192	1.2497885482659643	0.42571428571428643	425.71428571428646	7.015	29.863857142857192
		720	30	3.63	0.2	15	1827	0.6139758367501926	0.08847698795488651	0.05162127928336285	0.020959591148493852	2.462895335965193	0.4485714285714299	448.5714285714299	8.605	38.599571428571544
		751	31.291666666666668	4.52	0.2	10	1973	3.0911072165751454	0.44544400544456136	0.25989118686859125	0.17699210303172588	1.4683773028111073	0.4542857142857118	454.2857142857118	6.29	28.574571428571268
		771	32.125	4.83	0.2	10	2065	4.935143513650349	0.7111788560345836	0.41493232530138563	0.20493822456305102	2.024670244831403	0.43999999999999917	439.99999999999915	4.88	21.471999999999955
		793	33.041666666666664	4.8	0.2	10	2216	6.926795186353547	0.998185820329478	0.5823845295704917	0.21425359840682606	2.718201859390274	0.42571428571428643	425.71428571428646	2.81	11.962571428571449
		817	34.041666666666664	1.59	0.2	35	2378	7.964932040347393	1.1477865316742613	0.6696680173996261	0.22124012878965738	3.0268831475699813	0.42428571428571515	424.28571428571513	3.44	14.5954285714286
		842	35.083333333333336	1.476	0.2	35
		864	36	1.422	0.2	35
		890	37.083333333333336	1.372	0.2	40
		917	38.208333333333336	1.27	0.2	40
		939	39.125	1.326	0.2	40
		967	40.291666666666664	1.294	0.2	40
		989	41.208333333333336	1.268	0.2	40
		1009	42.041666666666664	1.363	0.2	40
		1035	43.125	1.7	0.2	30
		1060	44.166666666666664	1.748	0.2	30
		1084	45.166666666666664	1.738	0.2	30
		1108	46.166666666666664	1.33	0.2	40
		1132	47.166666666666664	1.246	0.2	40
		1155	48.125	1.318	0.2	40
		1184	49.333333333333336	1.41	0.2	35
		1204	50.166666666666664	1.46	0.2	35
		1227	51.125	1.53	0.2	35
		1250	52.083333333333336	1.606	0.1	35
		1278	53.25	1.574	0.1	35
		1330	55.416666666666664	1.548	0.1	35
		1348	56.166666666666664	1.7	0.1	35
		1376	57.333333333333336	1.778	0.1	35
		1396	58.166666666666664	1.782	0.1	35
		1423	59.291666666666664	2.036	0.075	35
		1445	60.208333333333336	2.03	0.075	35
		1471	61.291666666666664	1.972	0.075	35
		1496	62.333333333333336	1.974	0.075	35
		1515	63.125	1.972	0.075	35
		1563	65.125	1.914	0.075	35
		1827	76.125	2.05	0.0522	35
		1899	79.125	1.88	0.075	35
		1973	82.20833333333333	1.76	0.075	35
		2045	85.20833333333333	1.84	0.1	35
		2065	86.04166666666667	1.65	0.1	35
		2216	92.33333333333333	1.692	0.125	35
		2378	99.08333333333333	1.67	0.15	35

Experiment	1 L Continuous - 100 days

						STANDARDS
	Experiment	Timepoint[h]	Peak area	BA-Na [mM]
	CF6	-1	14553.03	22.139650658848357
		0	15705.892	23.893509610411105
		1	14910.882	22.684054007674696		Agilent infinity 2				Run Date	Agilent infinity 2
		2	14616.676	22.23647587021898
		3	13979.13	21.2665716153012		BA standard					BA standard
		5	9078.971	13.8119172627154
		8	2215.032	3.3697473775681215		Concentration [mM]	Peak area				Concentration [mM]	Peak area
		24	0	0		100	38930.897				100	36398.443	Peak not sharp
		48	0	0		50	26488.097	641.9249867451587	172.10082958946623		50	25804.735		657.3287688456737	162.12088536018564
		79	0	0		25	15972.812	10.006308583743312	102.12568016315453		25	16300.025		12.246680773997813	124.99120862787477
		99	0	0		12.5	8583.559	0.9985442132180414	228.89247830466408		12.5	8866.612		0.997921647497018	280.1405823043074
		122	0	0		6.25	4404.592	4115.482674768923	6		6.25	4564.418		2880.9020011722055	6
		144	0	0		3.125	2238.151	2.1561740784346524E8	314350.59974670666		3.125	2088.798		2.2608957597968706E8	470872.47512277856
		169	0	0		1.5625	1133.563				1.5625	1179.658
		192	0	0		0.78125	574.736				0.78125	595.297
		263	10313.687	15.690301413843422		0.390625	291.076				0.390625	293.601
		335	1464.561	2.2280493415167566		0.1953125	149.191				0.1953125	146.612
		434	3466.651	5.273846209081361
		505	6152.548	9.359924591772032
		578	4707.813	7.162036715871874
		650	3175.93	4.831569832327443
		720	0	0
		793	0	0
		864	5449.217	8.289941046246566
		939	6460.915	9.829045981250175
		1009	584.131	0.8886435525113154
		1084	0	0
		1155	5890.543	8.961334114677463
		1227	5231.63	7.958924057488429
		1250	2019.164	3.0717716917317484
		1278	1374.508	2.0910509321970983
		1330	1319.7	2.0076710468185786
		1348	1593.299	2.423899576589372
		1396	1885.056	2.867752029121499
		1563	1342.561	2.042449684237173
		1827	403.584	0.6139758367501926
		1827	404.321	0.6298570705300465	crosscheck
		1973	1984.259	3.0911072165751454
		2065	3167.992	4.935143513650349
		2216	4446.483	6.926795186353547
		2378	5112.889	7.964932040347393
		2570	5564.174	8.667950305721074

Experiments	Continuous fermentation
Date	July - Sep, 2023

Retention times [min]
Internal standard	9.82

C4	6.03
C6	10.32
C8	17.1
C10	24.22
C12	32.96



			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
CF6-24	9.2	0.85	130.1	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0										0
CF6-48	9.9	0.85	152.9	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0										0
CF6-79	13.3	0.85	157.4	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0										0
CF6-99	10	0.85	137	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0										0
CF6-122	11.1	0.85	131	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0										0
CF6-144	10.4	0.85	128.6	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0										0
CF6-169	10.9	0.85	134.4	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0										0
CF6-192	14.9	0.85	137.5	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0										0
CF6-217	10.6	0.85	144.2	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0										0
CF6-242	9.9	0.85	141.6	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0										0
CF6-263	8.9	0.85	149.6	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0										0
CF6-290	10.7	0.85	146.6	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0										0
CF6-314	9.4	0.85	164	0	0	0	0	0	0	84.7	0	0	0	0	0	0	0	0	0.0952118245186136	0	0	0.0952118245186136	0	0	0	0	0	0	100	0	0	1.1916373531741375
CF6-335	12.7	0.85	164.3	0	0	0	0	0	0	123.9	0	0	0	0	0	0	0	0	0.13902249171925551	0	0	0.13902249171925551	0	0	0	0	0	0	100	0	0	1.287841516621172
CF6-363	11.2	0.85	166	0	0	0	0	0	0	95.7	0	0	0	0	0	0	0	0	0.10628088814204183	0	0	0.10628088814204183	0	0	0	0	0	0	100	0	0	1.1163958838449772
CF6-416	10.3	0.85	159.3	0	0	0	0	0	0	112.5	0	0	0	0	0	0	0	0	0.13019313113291703	0	0	0.13019313113291703	0	0	0	0	0	0	100	0	0	1.4870717433799774
CF6-434	10.1	0.85	155.8	0	0	0	0	0	0	122.5	0	0	0	0	0	0	0	0	0.14495058104182149	0	0	0.14495058104182149	0	0	0	0	0	0	100	0	0	1.68841678557742
CF6-462	9	0.85	168.3	0	0	0	0	0	0	144.9	0	0	0	0	0	0	0	0	0.15872143990993523	0	0	0.15872143990993523	0	0	0	0	0	0	100	0	0	2.07479006418216
CF6-482	8.7	0.85	171.5	0	0	0	0	27.3	0	156	0	0	0	0	0	0	0.031056479999999997	0	0.16769178947368418	0	0	0.19874826947368418	0	0	0	0	15.626037943496215	0	84.37396205650379	0	0	2.6876033735454254
CF6-505	9.1	0.85	163.7	0	0	0	0	0	0	146.4	0	0	0	0	0	0	0	0	0.1648707913706073	0	0	0.1648707913706073	0	0	0	0	0	0	100	0	0	2.1314905154571084
CF6-529	11.3	0.85	166.1	0	0	0	0	0	0	183.8	0	0	0	0	0	0	0	0	0.20399860603948158	0	0	0.20399860603948158	0	0	0	0	0	0	100	0	0	2.1238792924464502
CF6-553	11.5	0.85	169	0	0	0	0	0	0	165.5	0	0	0	0	0	0	0	0	0.18053550233572094	0	0	0.18053550233572094	0	0	0	0	0	0	100	0	0	1.8469105098283474
CF6-578	12	0.85	172.3	0	0	0	0	31.3	0	193.1	0	0	0	0	0	0	0.0354415549622751	0	0.20660856510981457	0	0	0.24205012007208968	0	0	0	0	14.642238124781578	0	85.35776187521842	0	0	2.3730403928636243
CF6-606	13	0.85	168.2	0	0	0	0	32.6	0	189.2	0	0	0	0	0	0	0.03781336409036861	0	0.20737025696226297	0	0	0.24518362105263158	0	0	0	0	15.422467425852856	0	84.57753257414714	0	0	2.218856299118838
CF6-628	12.9	0.85	167	0	0	0	0	28.3	0	183.8	0	0	0	0	0	0	0.033061585149700604	0	0.20289921235423886	0	0	0.23596079750393945	0	0	0	0	14.011473727600293	0	85.98852627239971	0	0	2.1519452576738662
CF6-650	13.8	0.85	168.9	0	0	0	0	28.5	0	186.6	0	0	0	0	0	0	0.03292068916518649	0	0.20367293185005136	0	0	0.23659362101523784	0	0	0	0	13.91444495583684	0	86.08555504416316	0	0	2.016995916583443
CF6-677	11.9	0.85	162.5	0	0	0	0	29.3	0	190.6	0	0	0	0	0	0	0.03517774227692307	0	0.21623244359514165	0	0	0.2514101858720647	0	0	0	0	13.99217066520288	0	86.00782933479712	0	0	2.4855183971533834
CF6-700	12.9	0.85	164	0	0	0	0	31.2	0	210.4	0	0	0	0	0	0	0.03711628097560976	0	0.23651201745827982	0	0	0.2736282984338896	0	0	0	0	13.564489195030133	0	86.43551080496987	0	0	2.49547011795613
CF6-720	9.4	0.85	148.2	0	0	0	0	0	0	124.7	0	0	0	0	0	0	0	0	0.15512063541444707	0	0	0.15512063541444707	0	0	0	0	0	0	100	0	0	1.9414347361007143
CF6-751	11.1	0.85	149.8	0	0	0	0	0	0	38	0	0	0	0	0	0	0	0	0.04676523364485981	0	0	0.04676523364485981	0	0	0	0	0	0	100	0	0	0.49565695437053325
CF6-771	10.9	0.85	157.3	0	0	0	0	0	0	37.2	0	0	0	0	0	0	0	0	0.043597897145916276	0	0	0.043597897145916276	0	0	0	0	0	0	100	0	0	0.4705655385419997
CF6-793	13.2	0.85	161.7	0	0	0	0	0	0	52.8	0	0	0	0	0	0	0	0	0.06019705263157894	0	0	0.06019705263157894	0	0	0	0	0	0	100	0	0	0.5365156206023078
CF6-817	11	0.85	167.5	0	0	0	0	0	0	90.8	0	0	0	0	0	0	0	0	0.09993609200314216	0	0	0.09993609200314216	0	0	0	0	0	0	100	0	0	1.068835208589756
CF6-842	8.6	0.85	167	0	0	0	0	0	0	63.3	0	0	0	0	0	0	0	0	0.06987769391742829	0	0	0.06987769391742829	0	0	0	0	0	0	100	0	0	0.9559192054367756
CF6-864	10.2	0.85	170.4	0	0	0	0	0	0	81.6	0	0	0	0	0	0	0	0	0.08828194514455151	0	0	0.08828194514455151	0	0	0	0	0	0	100	0	0	1.018246195438887
CF6-890	12.9	0.85	188.8	0	0	0	0	0	0	144.5	0	0	0	0	0	0	0	0	0.1410968058652988	0	0	0.1410968058652988	0	0	0	0	0	0	100	0	0	1.2867925751509237
CF6-917	12.9	0.85	196.9	0	0	0	0	0	0	133.7	0	0	0	0	0	0	0	0	0.1251805964021277	0	0	0.1251805964021277	0	0	0	0	0	0	100	0	0	1.141637906084156
CF6-939	13	0.85	196.5	0	0	0	0	0	0	135.4	0	0	0	0	0	0	0	0	0.12703033250301324	0	0	0.12703033250301324	0	0	0	0	0	0	100	0	0	1.1495957692580385
CF6-967	13.5	0.85	170.3	0	0	0	0	28.7	0	203.4	0	0	0	0	0	0	0.032879178391074564	0	0.22018494743023148	0	0	0.25306412582130605	0	0	0	0	12.992429600350091	0	87.00757039964991	0	0	2.2053518590092036
CF6-989	13.5	0.85	168.3	0	0	0	0	0	0	191.2	0	0	0	0	0	0	0	0	0.20943781442912088	0	0	0.20943781442912088	0	0	0	0	0	0	100	0	0	1.8251661388158682
CF6-1009	14	0.85	164.7	0	0	0	0	29.2	0	204	0	0	0	0	0	0	0.034589394535519125	0	0.22834310037388553	0	0	0.2629324949094046	0	0	0	0	13.15523763901345	0	86.84476236098656	0	0	2.2095167639445767
CF6-1035	12	0.85	163.2	0	0	0	0	0	0	167.9	0	0	0	0	0	0	0	0	0.18966267298761608	0	0	0.18966267298761608	0	0	0	0	0	0	100	0	0	1.8594379704668242
CF6-1060	12.8	0.85	160.6	0	0	0	0	0	0	173.3	0	0	0	0	0	0	0	0	0.19893186170282495	0	0	0.19893186170282495	0	0	0	0	0	0	100	0	0	1.8284178465333176
CF6-1084	12.7	0.85	158.5	0	0	0	0	0	0	165.6	0	0	0	0	0	0	0	0	0.19261157881454422	0	0	0.19261157881454422	0	0	0	0	0	0	100	0	0	1.7842665939281541
CF6-1108	11	0.85	158.8	0	0	0	0	0	0	81.8	0	0	0	0	0	0	0	0	0.09496293543682882	0	0	0.09496293543682882	0	0	0	0	0	0	100	0	0	1.0156463683083297
CF6-1132	12.5	0.85	173.7	0	0	0	0	0	0	109.9	0	0	0	0	0	0	0	0	0.11664045341332605	0	0	0.11664045341332605	0	0	0	0	0	0	100	0	0	1.097792502713657
CF6-1155	12.7	0.85	171.1	0	0	0	0	0	0	116.1	0	0	0	0	0	0	0	0	0.1250931519271586	0	0	0.1250931519271586	0	0	0	0	0	0	100	0	0	1.158806409700404
CF6-1184	14.5	0.85	178.4	0	0	0	0	0	0	132.8	0	0	0	0	0	0	0	0	0.1372317337738966	0	0	0.1372317337738966	0	0	0	0	0	0	100	0	0	1.1134420590174168

Experiment	1 L Continuous - 100 days
Date	aug-oct, 2022



				N=OD/0.429397688926138
				20/10/2022						CF6 [100 x diluted samples]
				CF6 [undiluted samples]					N=OD/0.409410534471549
					OD at 635 nm	N-concentration [mg/l] - diluted	Dilution factor	N-concentration [mg/l]	2022-03-08T0:0:0+0000
				-1	3.328	7.7503910380208385	1	7.7503910380208385	CF6
				0	3.216	7.489560570395138	1	7.489560570395138		OD at 635 nm	N-concentration of diluted sample [mg/l]	N [mg/l]
				1	3.342	7.782994846474052	1	7.782994846474052	-1	1.019	2.488944260594772	248.8944260594772
				2	3.341	7.780666003013108	1	7.780666003013108	0	0.997	2.435208466941107	243.5208466941107
				3	2.997	6.979543852448454	1	6.979543852448454	1	0.971	2.371702528986775	237.1702528986775
				5	2.408	5.607855053952578	1	5.607855053952578	2	0.943	2.30331151888211	230.331151888211
				8	3.039	7.077355277808092	1	7.077355277808092	3	0.881	2.1518742822217805	215.18742822217806
				24	0.497	1.1574352000890495	1	1.1574352000890495	5	0.656	1.602303665309294	160.2303665309294
				48	0.501	1.1667505739328246	1	1.1667505739328246	8	0.101	0.2466961435918273	24.669614359182727
				79	0.288	0.6707069167518033	1	0.6707069167518033	24	0.007	0.017097752526166247	1.7097752526166248
				99	0.279	0.6497473256033096	1	0.6497473256033096
				122	0.2	0.4657686921887524	1	0.4657686921887524
				144	0.671	1.5626539622932643	1	1.5626539622932643
				169	0.69	1.6069019880511954	1	1.6069019880511954
				192	0.643	1.4974463453868387	1	1.4974463453868387
				263	0.103	0.23987087647720745	1	0.23987087647720745
				335	0.125	0.29110543261797023	1	0.29110543261797023
				434	0.121	0.28179005877419516	1	0.28179005877419516
				505	0.107	0.2491862503209825	1	0.2491862503209825
				578	0.071	0.16534788572700707	1	0.16534788572700707
				650	0.051	0.11877101650813184	1	0.11877101650813184
				720	-0.024	-0.05589224306265028	1	-0.05589224306265028
				793	0.022	0.051234556140762756	1	0.051234556140762756
				864	0.033	0.07685183421114414	1	0.07685183421114414
				939	0.055	0.1280863903519069	1	0.1280863903519069
				1009	0.012	0.02794612153132514	1	0.02794612153132514
				1084	0.076	0.17699210303172588	1	0.17699210303172588
				1155	0.004	0.009315373843775046	1	0.009315373843775046
				1227	0.033	0.07685183421114414	1	0.07685183421114414
				1396	-0.022	-0.051234556140762756	1	-0.051234556140762756
				1563	0.059	0.13740176419568192	1	0.13740176419568192
				1827	0.009	0.020959591148493852	1	0.020959591148493852
				1973	0.076	0.17699210303172588	1	0.17699210303172588
				2065	0.088	0.20493822456305102	1	0.20493822456305102
				2216	0.092	0.21425359840682606	1	0.21425359840682606
				2378	0.095	0.22124012878965738	1	0.22124012878965738







	NanoPlateReader
	2022-03-08T0:0:0+0000
	N-std
													0.40941053447154946	0.001100543099846793	slope	intercept
	Sample	N-concentration in well [g/l]	N-concentration in well [mg/l]	OD at 635nm									0.0022685222720699814	0.0010845130724574821	se(slope)	se(intercept)
	2x	0.00130925	1.30925	0.535									0.9997544435203787	0.0028705513616219067	r2	se(y)
	1x	6.54625E-4	0.654625	0.273									32571.06292001101	8	F	df
	0.5	3.273125E-4	0.3273125	0.136									0.26838767947904235	6.592052095767506E-5	ss reg	ss resid
	0.25	1.6365625E-4	0.16365625	0.07
	0.125	8.1828125E-5	0.081828125	0.029
	0.0625	4.09140625E-5	0.0409140625	0.02
	0.03125	2.045703125E-5	0.02045703125	0.009
	0.015625	1.0228515625E-5	0.010228515625	0.007
	0.0078125	5.1142578125E-6	0.0051142578125	0.002
	0.00390625	2.55712890625E-6	0.00255712890625	0.001






	NanoPlateReader
	21/10/2022
	N-std

	Sample	N-concentration in well [g/l]	N-concentration in well [mg/l]	OD at 635nm
	10x	0.00654625	6.54625	3.321
	5	0.003273125	3.273125	1.404
	2.5	0.0016365625	1.6365625	0.702
	1.25	8.1828125E-4	0.81828125	0.355
	0.625	4.09140625E-4	0.409140625	0.178										0.42939768892613767	-2.4554608830362756E-4	slope	intercept
	0.3125	2.045703125E-4	0.2045703125	0.092										0.0012897883571468163	0.001723463558177953	se(slope)	se(intercept)
	0.15625	1.0228515625E-4	0.10228515625	0.044										0.999945869100962	0.0038627683504178193	r2	se(y)
	0.078125	5.1142578125E-5	0.051142578125	0.018										110836.42284195931	6	F	df
	0.0390625	2.55712890625E-5	0.0255712890625	0.005										1.6537879741240258	8.952587597393761E-5	ss reg	ss resid
	0.01953125	1.278564453125E-5	0.01278564453125	-0.002

Exp	G45	WT P. putida CA3_Benzoate
Aim	To understand relation between growth, N and C concentrations and PHA accumulation
Plan	Culture the bug under N-lim with BA-Na as C source. Harvest the flasks every hour for 24 h. Single flask sufficient [with BI and BII crosscheck].																	Volumetric productivity
											Time [h]	C [mg/L]	N [mg/L]	C/N [g/g]	CDW [mg/L]	PHA [mg/L]	CDW - PHA [mg/L]	Biomass [mg/l/h]	PHA [mg/l/h]
Agar plate	MSM BA										1	1880.5486432508806	76.85257383015215	24.46955969759689	0	0	0	0	0
Inoculum	10 ml MSM full N + BA-na										2	1977.7930205871521	77.06390079741075	25.6643253212223	0	0	0	0	0
Batch I					Batch II						3	1992.2956492502447	74.10532325579041	26.88464960031831	5.999999999999339	0	5.999999999999339	1.9999999999997797	0
T23	OD 540nm	Vol[ml] for 0.005x50 ml			T22	OD 540nm	Vol[ml] for 0.005x50 ml				4	1954.1870051927342	70.93541874691145	27.548818907590082	5.999999999999339	0	5.999999999999339	1.4999999999998348	0
1	1.24	0.20161290322580647			1.35	0.18518518518518517					5	1918.6798152566785	61.21437825301598	31.34361354331238	-7.999999999999119	0	-7.999999999999119	-1.5999999999998238	0
2	1.214	0.20593080724876442			1.15	0.2173913043478261					6	1857.217940340769	52.97262652993068	35.05995571677762	67.99999999999696	0	67.99999999999696	11.333333333332826	0
											7	1713.7376996493126	36.4891230837601	46.965713473449604	148.00000000000148	3.596400000000036	144.40360000000143	21.142857142857356	0.5137714285714338
											8	1555.9346609517352	19.37163873581373	80.32023940623918	240.00000000000023	2.5920000000000027	237.40800000000021	30.00000000000003	0.32400000000000034
											9	1405.9357226656357	12.186521849021426	115.36808780091195	304.00000000000205	4.347200000000029	299.652800000002	33.777777777778006	0.48302222222222546
											10	1238.190786045233	3.944770125936136	313.8816069165493	369.9999999999992		369.9999999999992	36.99999999999992	0
											11	1155.5439952936147	0.2613410168808884	4421.594471028931	424.00000000000216	5.681600000000029	418.31840000000216	38.54545454545474	0.5165090909090936
											12	977.8405267420583	0.011975195515743732	81655.49576677024	496.0000000000031	22.07200000000014	473.928000000003	41.33333333333359	1.839333333333345
											13	872.1748879923806	0.7431665022304901	1173.5928427542058	529.9999999999993	69.27099999999992	460.7289999999994	40.76923076923072	5.3285384615384555
											14	854.7810853147383	0.6903347604158407	1238.2124359489578	546.000000000002	75.78480000000029	470.21520000000174	39.00000000000015	5.41320000000002
											15	739.1988270958373	0.7326001538675602	1009.00719607202	607.9999999999997	113.02719999999994	494.9727999999997	40.53333333333331	7.535146666666662
											16	629.7266199677566	0.7114674571417005	885.1095206766937	626.0000000000022	132.58680000000044	493.4132000000017	39.125000000000135	8.286675000000027
											17	520.699863724132	0.519159916936377	1002.9662282035224	662.0000000000026	165.50000000000065	496.50000000000193	38.94117647058839	9.735294117647097
											18	334.0882943185572	0.4346291300329381	768.6744197132817	758.0000000000009	222.85200000000026	535.1480000000006	42.111111111111164	12.38066666666668
											19	162.2297805361098	0.16201734226934775	1001.311206959616	798.0000000000009	265.4148000000003	532.5852000000007	42.00000000000005	13.969200000000015
											20	94.17206290031548	0.033107892241603466	2844.3992209802655	828.000000000002	292.0356000000007	535.9644000000013	41.400000000000105	14.601780000000037
											21	2.6383894785312307	0.018315004533501652	144.05617392587104	851.9999999999994	294.1103999999998	557.8895999999996	40.57142857142854	14.005257142857133
											22	0	0		859.9999999999985	303.2359999999995	556.763999999999	39.09090909090902	13.783454545454521
											23	0	0		853.9999999999992	331.1811999999997	522.8187999999996	37.13043478260866	14.39918260869564
											24	0	0		854.0000000000036	314.35740000000135	539.6426000000023	35.583333333333485	13.098225000000056
										Crosscheck	12	1020.2968028662071	-0.007044231537530002	-144841.46317881614	468.00000000000176	27.658800000000102	440.34120000000166	39.00000000000015	2.3049000000000084
Sample [T-h]	OD 540 nm	Wi[g]	Wf[g]	CDW [g/l]	PHA [%CDW]	PHA [mg/l]	C [mg/l]	N [mg/l]	CDW [mg/l]
B1 control							1982.4084994808165
B2 control							1992.313200093678
1		1.0427	1.0427	0		0	1880.5486432508806	76.85257383015215	0
2		1.0431	1.0421	-0.019999999999997797		0	1977.7930205871521	77.06390079741075	-19.999999999997797
3		1.0551	1.0554	0.005999999999999339		0	1992.2956492502447	74.10532325579041	5.999999999999339
4		1.061	1.0613	0.005999999999999339		0	1954.1870051927342	70.93541874691145	5.999999999999339
5		1.0584	1.058	-0.007999999999999119		0	1918.6798152566785	61.21437825301598	-7.999999999999119
6		1.0535	1.0569	0.06799999999999695		0	1857.217940340769	52.97262652993068	67.99999999999696
7		1.0635	1.0709	0.14800000000000146	2.43	3.596400000000036	1713.7376996493126	36.4891230837601	148.00000000000148
8		1.0689	1.0809	0.2400000000000002	1.08	2.5920000000000027	1555.9346609517352	19.37163873581373	240.00000000000023
9		1.0568	1.072	0.30400000000000205	1.43	4.347200000000029	1405.9357226656357	12.186521849021426	304.00000000000205
10		1.0578	1.0763	0.3699999999999992			1238.190786045233	3.944770125936136	369.9999999999992
11		1.053	1.0742	0.42400000000000215	1.34	5.681600000000029	1155.5439952936147	0.2613410168808884	424.00000000000216
12		1.0571	1.0819	0.4960000000000031	4.45	22.07200000000014	977.8405267420583	0.011975195515743732	496.0000000000031
13		1.0442	1.0707	0.5299999999999994	13.07	69.27099999999992	872.1748879923806	0.7431665022304901	529.9999999999993
14		1.0606	1.0879	0.546000000000002	13.88	75.78480000000029	854.7810853147383	0.6903347604158407	546.000000000002
15		1.0539	1.0843	0.6079999999999997	18.59	113.02719999999994	739.1988270958373	0.7326001538675602	607.9999999999997
16		1.0553	1.0866	0.6260000000000021	21.18	132.58680000000044	629.7266199677566	0.7114674571417005	626.0000000000022
17		1.0547	1.0878	0.6620000000000026	25	165.50000000000065	520.699863724132	0.519159916936377	662.0000000000026
18		1.0573	1.0952	0.7580000000000009	29.4	222.85200000000026	334.0882943185572	0.4346291300329381	758.0000000000009
19		1.0539	1.0938	0.7980000000000009	33.26	265.4148000000003	162.2297805361098	0.16201734226934775	798.0000000000009
20		1.0532	1.0946	0.8280000000000021	35.27	292.0356000000007	94.17206290031548	0.033107892241603466	828.000000000002
21		1.0505	1.0931	0.8519999999999994	34.52	294.1103999999998	2.6383894785312307	0.018315004533501652	851.9999999999994
22		1.0608	1.1038	0.8599999999999985	35.26	303.2359999999995	0	-7.044225197720958E-4	859.9999999999985
23		1.0546	1.0973	0.8539999999999992	38.78	331.1811999999997	0	-0.009157501210115975	853.9999999999992
24		1.0453	1.088	0.8540000000000036	36.81	314.35740000000135	0	-0.004930961864944043	854.0000000000036
12crosscheck		1.0543	1.0777	0.46800000000000175	5.91	27.658800000000102	1020.2968028662071	-0.007044231537530002	468.00000000000176

Experiments	C/N ratio study - shake flasks [G45]
Date	March, 2023

Retention times [min]
Internal standard	9.17

C4	5.56
C6	9.64
C8	16.28
C10	23.35
C12	31.45



			GC - peak area data										Mass of PHA monomer										PHA monomer distribution
Sample name [hour]	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
T7	9.2	0.85	299.2	0	0	0	0	0	0	308.4	0	0	0	0	0	0	0	0	0.1900220965381368	0	0	0.1900220965381368	0	0	0	0	0	0	100	0	0	2.4299500836078876
T8	9.7	0.85	295.3	0	0	0	0	0	0	142.2	0	0	0	0	0	0	0	0	0.08877434459158393	0	0	0.08877434459158393	0	0	0	0	0	0	100	0	0	1.076705210328489
T9	10.9	0.85	337.5	0	0	0	0	12.1	0	188	0	42.8	0	0	0	0	0.006994638933333333	0	0.10269171274853797	0	0.022490089120772945	0.13217644080264426	0	0	0	0	5.291895356584153	0	77.69290209733319	0	17.015202546082644	1.4266210556140775
T11	6.4	0.85	180.8	0	0	0	0	0	0	71.6	0	0	0	0	0	0	0	0	0.07300723847228689	0	0	0.07300723847228689	0	0	0	0	0	0	100	0	0	1.3420448248582149
T12	9.7	0.85	221	0	0	0	0	88.8	0	345.6	0	0	0	0	0	0	0.07839247927601808	0	0.2882921289830912	0	0	0.3666846082591093	0	0	0	0	21.378720979917386	0	78.6212790200826	0	0	4.447357286344565
T13	10.8	0.85	202.5	6.9	0	32.9	0	275.1	0	899.4	0	82.6	0.008635324444444445	0	0.034712801077441076	0	0.2650447893333333	0	0.8188025394152045	0	0.07233961687600644	1.1995350711464299	0.7198892847869316	0	2.893854620212568	0	22.095626523034667	0	68.25999165098621	0	6.030637920979618	13.066830840375053
T14	8.2	0.85	230.2	0	0	33.5	0	262.5	0	822.5	0	71.3	0	0	0.03109268857120291	0	0.22247319721980888	0	0.6586912776075724	0	0.054929468288444826	0.967186631687029	0	0	3.2147558240097927	0	23.00209596898138	0	68.10384428687158	0	5.6793039201372455	13.876422262367706
T15	10	0.85	204.4	8.9	0	42.9	0	373.2	0	1199.9	0	98.9	0.011034780821917809	0	0.04484305479452054	0	0.35621684383561636	0	1.0822198291276133	0	0.08580973972602739	1.5801242483056952	0.6983489326076712	0	2.8379448541849777	0	22.543597075835876	0	68.48954000218875	0	5.430569135182742	18.58969703889053
T16	12	0.85	202.9	10.4	0	60	0	513.5	0	1625.1	0	130.4	0.012989902414982749	0	0.06318121779649626	0	0.49375568457368146	0	1.476554115742782	0	0.11397687067949516	2.160457791207438	0.6012569404433005	0	2.924436573286882	0	22.85421573997662	0	68.3445018806669	0	5.275588865626285	21.18095873732782
T17	13.2	0.85	180.3	10.8	0	72.2	0	594.8	0	1875.9	0	144.9	0.015180379367720466	0	0.08555792830131598	0	0.6436191254575706	0	1.918073662142044	0	0.14252591680532445	2.8049570120739755	0.5411982893989575	0	3.0502402686754455	0	22.945775022116326	0	68.38157069379922	0	5.08121572601005	24.999616863404423
T18	12.7	0.85	239.4	16.7	0	112.7	0	921.8	0	2797.2	0	202.3	0.01767856140350877	0	0.10058160446570974	0	0.7512184842105262	0	2.154024797783933	0	0.1498624820747521	3.17336592993843	0.5570918007508757	0	3.169555818217953	0	23.672608227223936	0	67.87823545536476	0	4.72250869844247	29.396627419531544
T19	12.9	0.85	192.7	14.3	0	106.4	0	856.7	0	2584.8	0	184.1	0.01880654073689673	0	0.11797193489644762	0	0.867362736274001	0	2.4728430657963014	0	0.1694311499289276	3.646415427632574	0.5157542005329555	0	3.23528509676805	0	23.78672297459903	0	67.81572519294075	0	4.64651253515923	33.255042659667794
T20	13.8	0.85	185.5	14.8	0	111	0	936.4	0	2829.1	0	201	0.020219592452830188	0	0.1278491526586621	0	0.9848525162264151	0	2.811614083018867	0	0.1921645742411813	4.136699918597956	0.48878557426720903	0	3.0906073724098837	0	23.807685730325087	0	67.96756202639426	0	4.645359296603542	35.26598396076689
T21	9.7	0.85	186.9	10.3	0	88.8	0	663.9	0	1941.3	0	129.3	0.013966337078651686	0	0.10151318488253319	0	0.6930220853932583	0	1.914849643997634	0	0.12269035075720568	2.8460416021092825	0.49072849350834635	0	3.56682013387643	0	24.350384930411412	0	67.2811543787162	0	4.3109120634876295	34.51839420387245
T22	13.2	0.85	244.1	17.8	0	152.4	0	1202.4	0	3531.9	0	242.2	0.018480206472757065	0	0.1333939020520651	0	0.9610254656288406	0	2.6674233252980875	0	0.17596550651016155	3.956288405961912	0.46710968909416206	0	3.371693070986628	0	24.291087176067027	0	67.4223679264238	0	4.4477421374283805	35.261037486291556
T23	11.9	0.85	196	14.2	0	120	0	958.2	0	2818.6	0	186.7	0.018360599999999998	0	0.1308109090909091	0	0.95379228	0	2.651115821052631	0	0.16893103043478258	3.9230106405783225	0.4680232016218369	0	3.334452059289475	0	24.312763012526368	0	67.57860388219108	0	4.306157844371258	38.784089377936944
T24	13.5	0.85	235.7	18.2	0	152.4	0	1222.4	0	3674.9	0	239.1	0.019568899448451423	0	0.13814786377135804	0	1.0118298012728044	0	2.874334240314405	0	0.179904148530741	4.22378495333776	0.46330245655587887	0	3.2707125314746315	0	23.955523599117576	0	68.05115014302565	0	4.2593112698262585	36.808583471353025
T12 crosscheck	9.5	0.85	222.4	0	0	11	0	105.5	0	405.6	0	47.7	0	0	0.010567607913669064	0	0.09254892625899279	0	0.3362129897765997	0	0.03803687073819205	0.4773663946874536	0	0	2.21373101066069	0	19.387398712803694	0	70.43080399422095	0	7.968066282314647	5.911658138544317

Experiments	C/N ratio study - shake flasks [G45]						N=OD/0.473200374269469
Date	March, 2023						2023-03-03T0:0:0+0000
							G45
							Sample	OD at 635 nm	N-concentration in well [mg/l]	Dilution	N [mg/l]
							1	0.363666667	0.7685257383015216	100	76.85257383015215
							2	0.364666667	0.7706390079741076	100	77.06390079741075
							3	0.35066666700000004	0.7410532325579041	100	74.10532325579041
							4	0.33566666700000003	0.7093541874691145	100	70.93541874691145
							5	0.28966666700000004	0.6121437825301598	100	61.21437825301598
							6	0.250666667	0.5297262652993068	100	52.97262652993068
							7	0.172666667	0.364891230837601	100	36.4891230837601
							8	0.091666667	0.1937163873581373	100	19.37163873581373
							9	0.057666667000000005	0.12186521849021427	100	12.186521849021426
							10	0.018666666999999998	0.03944770125936136	100	3.944770125936136
							11	0.123666667	0.2613410168808884	1	0.2613410168808884
							12	0.005666667	0.011975195515743732	1	0.011975195515743732
							13	0.35166666700000004	0.7431665022304901	1	0.7431665022304901
							14	0.326666667	0.6903347604158407	1	0.6903347604158407
							15	0.34666666700000004	0.7326001538675602	1	0.7326001538675602
							16	0.33666666700000003	0.7114674571417005	1	0.7114674571417005
							17	0.245666667	0.519159916936377	1	0.519159916936377
							18	0.20566666700000003	0.4346291300329381	1	0.4346291300329381
							19	0.07666666700000001	0.16201734226934775	1	0.16201734226934775
							20	0.01566666700000001	0.033107892241603466	1	0.033107892241603466
							21	0.008666667000000003	0.018315004533501652	1	0.018315004533501652
							22	-3.3333299999999816E-4	-7.044225197720958E-4	1	-7.044225197720958E-4
							23	-0.004333332999999995	-0.009157501210115975	1	-0.009157501210115975
							24	-0.002333333	-0.004930961864944043	1	-0.004930961864944043
							12 cc	-0.003333332999999994	-0.007044231537530002	1	-0.007044231537530002






	NanoPlateReader

	2023-03-03T0:0:0+0000			BLANK	No colour	OD 0.16
	N-std

		N-concentration in well [mg/l]	OD at 635nm
		65.4625	3.212666667
		32.73125	3.046666667
		16.365625	3.446666667
		8.1828125	3.250666667
		4.09140625	1.921666667
		2.045703125	1.032666667									0.4732003742694691	0.018168603483346257	slope	intercept
		1.0228515625	0.538666667									0.008326755751416793	0.013908154016021605	se(slope)	se(intercept)
		0.51142578125	0.27266666700000003									0.9981455862724676	0.031172099282808032	r2	se(y)
		0.255712890625	0.13266666700000002									3229.5239345559253	6	F	df
		0.1278564453125	0.06366666700000001									3.1381276763578163	0.005830198642183446	ss reg	ss resid
		0.06392822265625	0.027666667000000006
		0.031964111328125	0.012666667000000006

Experiments	C/N ratio study - shake flasks [G45]
Date	March, 2023


Run Date	Agilent infinity 2

	BA-na standard
	RT = 5.60
	Concentration [mM]	Peak area
	100	38930.897
	50	26488.097					641.9249867451587	172.10082958946623
	25	15972.812					10.006308583743312	102.12568016315453
	12.5	8583.559					0.9985442132180414	228.89247830466408
	6.25	4404.592					4115.482674768923	6
	3.125	2238.151					2.1561740784346524E8	314350.59974670666
	1.5625	1133.563
	0.78125	574.736
	0.390625	291.076
	0.1953125	149.191







	G45	BA-Na in undiluted samples
		Agilent 2		RT	5.6

	Samples [h]	Peak area	BA[mM]	BA [g/l]	C [g/l]	C [mg/l]
	Control1	15135.611	23.578472562994122	3.3977757886902684	1.9824084994808164	1982.4084994808165
	Control2	15211.233	23.69627760252366	3.4147520839116723	1.992313200093678	1992.313200093678
	1	14357.915	22.366966545936055	3.2231917141021156	1.8805486432508807	1880.5486432508806
	2	15100.372	23.52357674183121	3.3898650263815866	1.9777930205871521	1977.7930205871521
	3	15211.099	23.696068855395882	3.414722002406824	1.9922956492502446	1992.2956492502447
	4	14920.141	23.242810297153095	3.349405177871247	1.9541870051927341	1954.1870051927342
	5	14649.045	22.8204930482533	3.288547150718542	1.9186798152566784	1918.6798152566785
	6	14179.786	22.089474627098184	3.183203741137984	1.857217940340769	1857.217940340769
	7	13084.32	20.382941932468746	2.937283847178409	1.7137376996493126	1713.7376996493126
	8	11879.5	18.506056003427194	2.666815200373876	1.5559346609517353	1555.9346609517352
	9	10734.264	16.72199088678584	2.4097224967402737	1.4059357226656357	1405.9357226656357
	10	9453.538	14.726857498929004	2.122213799883164	1.238190786045233	1238.190786045233
	11	8822.533	13.743868832028664	1.9805602180394908	1.1555439952936146	1155.5439952936147
	12	7465.774	11.630290142929471	1.6759829610468515	0.9778405267420582	977.8405267420583
	13	6659.021	10.373518713245318	1.4948759141722165	0.8721748879923806	872.1748879923806
	14	6526.22	10.166639404914905	1.4650635714452624	0.8547810853147383	854.7810853147383
	15	5643.754	8.791921174592048	1.266959800864587	0.7391988270958373	739.1988270958373
	16	4807.938	7.489874985395491	1.0793284347704173	0.6297266199677566	629.7266199677566
	17	3975.523	6.193126922927134	0.8924605552284147	0.520699863724132	520.699863724132
	18	2550.751	3.9735966039646384	0.5726151386143242	0.3340882943185572	334.0882943185572
	19	1238.618	1.929536939673638	0.2780559206916696	0.1622297805361098	162.2297805361098
	20	719	1.1200685438330025	0.16140747750905482	0.09417206290031548	94.17206290031548
	21	20.144	0.031380612999961054	0.004522103236359388	0.002638389478531231	2.6383894785312307
	22	0	0	0	0	0
	23	0	0	0	0	0
	24	0	0	0	0	0
	12-crosscheck [cc]	7789.926	12.135258791914945	1.7487514682089031	1.0202968028662072	1020.2968028662071