本研究の一部はBMWiとBMBFプロジェクト」Aptasens」（BMBF / DLR 01RB0805A）が資金を提供し、IGF-プロジェクト「S-ふるい」（490 ZBG / 1）によって資金を供給されました。英語のネイティブスピーカーとして原稿を読み取るためのAFM研究中およびエリックV.ジョンストンへの彼の貴重な助けのためのトビアス・J・ギュンターに感謝します。さらに、この論文の著者は、Manjaフォーゲル、ナンシーアンガー、カレンE. Viacavaとヘルムホルツ研究所のグループバイオテクノロジー（ICP-MS測定における支援のためのリソースエコロジー研究所から）アリーンリッターとサブリナGurlitに感謝したいと思います資源技術のためのフライベルク。

著者らは、開示することは何もありません。

この研究では、異なる分析方法の組み合わせを用いて調べたS層タンパク質の金の結合を研究しました。具体的には、金の結合は、例えば 、感覚面鉱業水またはプロセス溶液から金を回収するだけでなく、材料の建設のためだけでなく、非常に魅力的です。 Auの相互作用の研究のために金（Au（III）及びAu（0）-NPs）に懸濁し、SLP1の単層を再結晶化して、タンパク質が持っていたが単離?...

原因エレクトロニクス、触媒、バイオセンサ、または医療機器のようないくつかのアプリケーションのための金の使用の増加に、この貴金属の需要はここ数年の時間6-9かけて成長してきました。重鎖または貴金属、最も環境汚染は継続的なプロセスであるが、金だけでなく、他の多くの貴重な重金属は、鉱業活動を通じて、希薄濃度の工業排水を経て環境中に放出され、廃棄物処理7,8,10されています主に技術的な活動によって引き起こされます。これは、自然の生態系の重要な干渉につながり、潜在的に人間の健康9を脅かします。これらの負の結果を知ることは、工業廃水からの金属のリサイクルに汚染された生態系や改善点から金属を除去するための新しい技術の探索を推進しています。沈殿又はイオン交換のような十分に確立された物理化学的方法には、特に高で、それほど効果的ではありませんLYは、溶液7,8,11に希釈しました。バイオソープションは、いずれかの生きているか死んバイオマスで、廃水処理10,12のための魅力的な代替です。そのような生物学的材料の使用は、毒性化学物質の消費量を低減することができます。多くの微生物は、蓄積又は金属を固定するために記載されています。例えば、Lysinibacillusのスフェリカスの細胞が（L.スファエリカス ）JG-A12は、貴金属のための高い結合能力を示している、例えば、パラジウム（II）、白金（II）、金（III）、及びPbのような他の有毒金属（II）またはU（VI）4,13、クロム（VI）14のためのバチルス・メガテリウムの細胞、金のために白金（II）およびPd（II）15、及びクロレラ下品ためのサッカロマイセス・セレビシエの細胞（III）とU（VI）16 、17。金（III）、パラジウム（II）、およびPt（II）のような前の金属の結合はまた、 デスルホビブリオ18およびL.ためのdesulfuricansについて報告されていますスフェリカス JG-B53 19,20。それにもかかわらず、ではないアルリットルの微生物は、金属を大量に結合して、収着材料としての用途は限ら12,21です。また、容量の金属結合は、異なるパラメータ、 例えば、細胞組成物、使用されるバイオコンポーネント、または環境や実験条件（pH、イオン強度、温度など）に依存します。単離された細胞壁断片22,23の研究は、膜脂質、ペプチドグリカン、タンパク質、または他の構成要素と同様に、複雑な構築全細胞8,21のプロセスを結合金属を理解するのに役立ちます。 この研究に着目セル構成要素は、S層タンパク質です。 S層タンパク質は、多くの細菌及び古細菌の外側細胞エンベロープの一部であり、それらは、約15を構成する - これらの生物の全タンパク質質量の20％。環境への最初のインターフェイスとして、これらの細胞の化合物は、強く、細菌の吸着特性3に影響を与えます 。 40の範囲の分子量を有するS層タンパク質kDaでの数百の細胞内で生産され、それらは脂質膜または高分子の細胞壁成分に層を形成することができる場合に外部組み立てられます。一旦単離されると、ほぼ全てのS層タンパク質は自発的に界面で、または平面または管状の構造体3を形成する面に、懸濁液中で自己集合する固有の特性を有しています。タンパク質の単分子層の厚さは、細菌に依存し5の範囲内である- 25nmで24。一般に、形成されたS層タンパク質の構造は、35ナノメートル3,24に対して斜め（P1またはP2）、正方形（P4）、または六角形（P3またはP6）2.5の格子定数との対称性を有することができます。格子形成は、二価の陽イオンに依存して、主のCa 2+ 25,26、ラフ、Jらに多くのケースであるように思われます。 S層タンパク質ベースのエンジニアナノ構造における産業用アプリケーションに基づくナノコンポジット。 （編Tijana Z.グローブ＆Aitziber L. Cortajarena）（スプリンガー、2016（提出））。それにもかかわらず、特に例えば、Ca 2+およびMg 2+などの二価カチオンのモノマーの折り畳み、モノマー-モノマー相互作用、格子を形成し、異なる金属の役割の完全な反応カスケードは、まだ完全には理解されていません。 グラム陽性菌株L. 27（新系統発生的分類した後、バチルス・スファエリカスから改称） スフェリカス JG-B53はウラン鉱山廃棄物の山「ハーバーランド」（Johanngeorgenstadt、ザクセン、ドイツ）4,28,29から単離しました。その機能S層タンパク質（SLP1）は正方格子、116 kDaで30の分子量、および細菌細胞31生きに ​​≈10nmの厚さを有しています。以前の研究では、約10nmの厚さで閉じ、安定したタンパク質層のインビトロでの形成は10分未満19で達成されました。関連菌株L.スフェリカス JG-A12は、また「ハーバーランド &quot;パイルからの分離株は、高い金属結合能力を有し、その単離されたS層タンパク質は、Auのような貴金属のための高い化学的および機械的安定性と優れた吸着率を示した（III）、白金（II）、およびPd（II）4,32,33。貴金属のこの結合は、いくつかの金属について多かれ少なかれ特異的であり、ポリマーの外側及び内側タンパク質表面上及び細孔内の官能基、イオン強度、およびpH値の利用可能性に依存します。 、OH - 、PO 4 - - 、SO 4 - 、およびSO-タンパク質による金属相互作用に関連する官能基は、COOH-、NH 2です。原理的には、金属結合能力は、アプリケーション、 ラフ、Jらの広いスペクトルを開きます。 S層タンパク質ベースのエンジニアナノ構造における産業用アプリケーションに基づくナノコンポジット。 （編Tijana Z.グローブ＆Aitziber L. Cortajarena）（スプリンガー、2016年（提出））。 例えば、除去又は回復のためbiosorptiveコンポーネントとして溶解毒性または有価金属の、合成または触媒反応、およびバイオ感覚層3,5,18,33のような他の生体工学材料のため、定期的に構造化金属ナノ粒子（NPS）の定義された堆積のためのテンプレート。 Auのような規則的に配置されたNPアレイ（0）-NPsはCO酸化34-37ための分子エレクトロニクスやバイオセンサー、超高密度記憶装置、および触媒に至るまでの主要な用途のために使用することができます。このようなアプリケーションや、これらの材料のスマートなデザインの開発は、基礎となる金属結合メカニズムのより深い理解を必要とします。 このようなバイオベース材料の開発のための前提条件は、生体分子と技術面38,39との間の界面層の信頼できる実装です。例えば、高分子電解質は、S層タンパク質39の再結晶化のための界面層として使用されているレイヤー・バイ・レイヤー（のLbL）技術40,41に組み付け</SUP&gt;。このようなインタフェースは、再現可能かつ定量的な方法でタンパク質コーティングを実行するための比較的簡単な方法を提供しています。接着剤プロモーターと変更することなく、種々の実験を行うことで、コーティング速度、層の安定性、および生体分子19,42、ラフ、J.らと金属との相互作用に関する記述することが可能です。 S層タンパク質ベースのエンジニアナノ構造における産業用アプリケーションに基づくナノコンポジット。 （編Tijana Z.グローブ＆Aitziber L. Cortajarena）（スプリンガー、2016年（提出））。しかし、タンパク質吸着およびタンパク質 - 表面相互作用の複雑なメカニズムは完全に理解されていません。特に立体構造、パターンの向き、コーティング密度の情報がまだ不足しています。 消費モニタリング（QCM-D）技術で水晶マイクロバランスは、タンパク質吸着、コーティング速度を研究するためのツール、および対話プロとして近年注目を集めていますナノメートルスケール19,43-45上セス。この技術は、リアルタイムで大量の吸着の詳細な検出を可能にし、タンパク質格子19,20,42,46-48に機能性分子のタンパク質自己集合プロセスと結合するための指標として用いることができます。また、QCM-Dの測定は、天然の生物学的条件下でのタンパク質性の層と金属の相互作用の過程を研究する可能性を開きます。最近の研究において、Euのような選択された金属とS層タンパク質の相互作用（III）、金（III）、パラジウム（II）、およびPtは（II）QCM-D 19,20で検討されています。吸着したタンパク質層は、グラム陽性細菌の細胞壁の単純化モデルとしての役割を果たすことができます。この単一の成分の研究では、金属相互作用のより深い理解に寄与することができます。しかし、単にQCM-D実験は、タンパク質に金属の表面構造と影響に関する記述を許可していません。他の技術は、そのような情報を得るために必要です。 One POSイメージングバイオナノ構造のための任及び構造特性情報を取得するには、原子間力顕微鏡（AFM）です。 提示研究の目的は、特にL. SLP1に、S層タンパク質に金（Au（III）とAu（0）-NPs）の収着を調査することでしたスフェリカス JG-B53。 ICP-MSを使用して5.0とQCM-Dを使用して固定化されたS-層と - 実験は、2.0のpH範囲内のバッチスケールで懸濁タンパク質を用いて行きました。さらに、格子の安定性に対する金属塩溶液の影響は、その後のAFM研究で調査しました。これらの技術の組み合わせは、特定の金属親和性について細菌細胞全体のイベントを結合についての詳細を学ぶためのツールとして、in vitroでの金属相互作用プロセスのより良い理解に貢献しています。この知識だけでなく、環境保護のための金属の回収のために適用可能なフィルター材料の開発と再の保全のために重要です49だけでなく、様々な技術的なアプリケーションのための高度に秩序化された金属NPのアレイの開発のためのソース。

<table border="0" cellpadding="0" cellspacing="0" width="849">
	<tbody>
		<tr height="20">
			<td height="20" style="height:20px;width:293px;">equiment and software</td>
			<td style="width:219px;"></td>
			<td style="width:97px;"></td>
			<td style="width:241px;"></td>
		</tr>
		<tr height="36">
			<td height="36" style="height:36px;width:293px;">Bioreactor, Steam In Place 70L Pilot System</td>
			<td style="width:219px;">Applikon Biotechnology, Netherlands</td>
			<td style="width:97px;">Z6X</td>
			<td style="width:241px;">Including dO2, pH sensors of Applikon Biotechnology and BioXpert software V2</td>
		</tr>
		<tr height="17">
			<td height="17" style="height:17px;width:293px;">Noninvasive Biomass Monitor BugEye 2100</td>
			<td style="width:219px;">BugLab, Concord (CA), USA</td>
			<td style="width:97px;">Z9X</td>
			<td style="width:241px;">---</td>
		</tr>
		<tr height="34">
			<td height="34" style="height:34px;width:293px;">Spectrometer Ultrospec 1000</td>
			<td style="width:219px;">Amersham Pharmacia Biotech, Great Britain</td>
			<td style="width:97px;">80-2109-10</td>
			<td style="width:241px;">Company now GE Healthcare Life Sciences</td>
		</tr>
		<tr height="17">
			<td height="17" style="height:17px;width:293px;">MiniStar micro centrifuge</td>
			<td style="width:219px;">VWR, Germany</td>
			<td style="width:97px;">521-2844</td>
			<td style="width:241px;">For centrifugation of cultivation samples</td>
		</tr>
		<tr height="34">
			<td height="34" style="height:34px;width:293px;">Research system microscope BX-61</td>
			<td style="width:219px;">Olympus Germany LLC, Germany</td>
			<td style="width:97px;">037006</td>
			<td style="width:241px;">Microscope in combination with imaging software</td>
		</tr>
		<tr height="34">
			<td height="34" style="height:34px;width:293px;">Cell^P (version 3.1)</td>
			<td style="width:219px;">Olympus Soft Imaging Solutions LLC, M&#252;nster, Germany</td>
			<td style="width:97px;">---</td>
			<td style="width:241px;">together with microscope</td>
		</tr>
		<tr height="17">
			<td height="17" style="height:17px;width:293px;">Powerfuge Pilot Separation System Serie 9010-S</td>
			<td style="width:219px;">Carr Centritech, Florida, USA</td>
			<td style="width:97px;">9010PLT</td>
			<td style="width:241px;">For biomasse harvesting</td>
		</tr>
		<tr height="34">
			<td height="34" style="height:34px;width:293px;">T18 basic Ultra Turrax</td>
			<td style="width:219px;">IKA Labortechnik, Germany</td>
			<td style="width:97px;">431-2601</td>
			<td style="width:241px;">For flagella removal and sample homogenization</td>
		</tr>
		<tr height="17">
			<td height="17" style="height:17px;width:293px;">Sorvall Evolution RC Superspeed Centrifuge</td>
			<td style="width:219px;">Thermo Fisher Scientific, USA</td>
			<td style="width:97px;">728411</td>
			<td style="width:241px;">Used within protein isolation</td>
		</tr>
		<tr height="34">
			<td height="34" style="height:34px;width:293px;">Mobile high shear fluid processor, M-110EH-30 Pilot</td>
			<td style="width:219px;">Microfluidics, Massachusetts, USA</td>
			<td style="width:97px;">M110EH30K</td>
			<td style="width:241px;">Used for cell rupture</td>
		</tr>
		<tr height="34">
			<td height="34" style="height:34px;width:293px;">Alpha 1-4 LSC Freeze dryer</td>
			<td style="width:219px;">Martin Christ Freeze dryers LLC, Osterode, Germany</td>
			<td>102041</td>
			<td style="width:241px;">---</td>
		</tr>
		<tr height="34">
			<td height="34" style="height:34px;width:293px;">UV-VIS spectrophotometry (NanoDrop 2000c)</td>
			<td style="width:219px;">Thermo Fisher Scientific, USA</td>
			<td style="width:97px;">91-ND-2000C-L</td>
			<td style="width:241px;">For determination of protein concentration</td>
		</tr>
		<tr height="34">
			<td height="34" style="height:34px;width:293px;">Mini-PROTEAN vertical electrophoresis chamber</td>
			<td style="width:219px;">Bio-Rad Laboratories GmbH, Munich, Germany</td>
			<td style="width:97px;">165-3322</td>
			<td style="width:241px;">For SDS-PAGE</td>
		</tr>
		<tr height="34">
			<td height="34" style="height:34px;width:293px;">VersaDoc Imaging System 3000</td>
			<td style="width:219px;">Bio-Rad Laboratories GmbH, Munich, Germany</td>
			<td style="width:97px;">1708030</td>
			<td style="width:241px;">Used for imaging of SDS-PAGE gels</td>
		</tr>
		<tr height="17">
			<td height="17" style="height:17px;width:293px;">ICP-MS Elan 9000</td>
			<td style="width:219px;">PerkinElmer, Waltham (MA), USA</td>
			<td style="width:97px;">N8120536</td>
			<td style="width:241px;">For determination of metal concentration</td>
		</tr>
		<tr height="34">
			<td height="34" style="height:34px;width:293px;">Zetasizer Nano ZS</td>
			<td style="width:219px;">Malvern Instruments, Worcestershire United Kingdom</td>
			<td style="width:97px;">ZEN3600</td>
			<td style="width:241px;">For determination of nanoparticle size</td>
		</tr>
		<tr height="17">
			<td height="17" style="height:17px;width:293px;">Q-Sense E4 device&#160;</td>
			<td style="width:219px;">Q-Sense AB, Gothenburg, Sweden</td>
			<td rowspan="3" style="width:97px;">QS-E4</td>
			<td rowspan="3" style="width:241px;">ordered via LOT quantum design (software included with E4 platform)</td>
		</tr>
		<tr height="17">
			<td height="17" style="height:17px;width:293px;">Q-Soft 401 (data recording)</td>
			<td style="width:219px;">Q-Sense AB, Gothenburg, Sweden</td>
		</tr>
		<tr height="17">
			<td height="17" style="height:17px;width:293px;">Q-Tools 3 (data evaluation and modelling)</td>
			<td style="width:219px;">Q-Sense AB, Gothenburg, Sweden</td>
		</tr>
		<tr height="17">
			<td height="17" style="height:17px;width:293px;">QCM-D flow modules QFM&#160;401&#160;</td>
			<td style="width:219px;">Q-Sense AB, Gothenburg, Sweden</td>
			<td style="width:97px;">QS-QFM401</td>
			<td style="width:241px;">ordered via LOT quantum design</td>
		</tr>
		<tr height="19">
			<td height="19" style="height:19px;width:293px;">QSX&#160;303 SiO2 piezoelectric AT-cut quartz sensors</td>
			<td style="width:219px;">Q-Sense AB, Gothenburg, Sweden</td>
			<td style="width:97px;">QS-QSX303</td>
			<td style="width:241px;">ordered via LOT quantum design</td>
		</tr>
		<tr height="17">
			<td height="17" style="height:17px;width:293px;">Ozone cleaning chamber</td>
			<td style="width:219px;">Bioforce Nanoscience, Ames (IA), USA</td>
			<td style="width:97px;">QS-ESA006</td>
			<td style="width:241px;">ordered via LOT quantum design</td>
		</tr>
		<tr height="17">
			<td height="17" style="height:17px;width:293px;">Atomic Force Microscope MFP-3D Bio AFM</td>
			<td style="width:219px;">Asylum Research, Santa Barbara (CA), USA</td>
			<td>MFP-3DBio</td>
			<td style="width:241px;">AFM measurements and imaging software</td>
		</tr>
		<tr height="34">
			<td height="34" style="height:34px;width:293px;">Asylum Research AFM Software AR Version 120804+1223</td>
			<td style="width:219px;">Asylum Research, Santa Barbara (CA), USA</td>
			<td style="width:97px;">---</td>
			<td style="width:241px;">imaging software included in Cat. No. MFP-3DBio</td>
		</tr>
		<tr height="34">
			<td height="34" style="height:34px;width:293px;">Igor Version Pro 6.3.2.3 Software</td>
			<td style="width:219px;">WaveMetrics, Inc., USA</td>
			<td style="width:97px;">---</td>
			<td style="width:241px;">imaging software included in Cat. No. MFP-3DBio</td>
		</tr>
		<tr height="17">
			<td height="17" style="height:17px;width:293px;">BioHeater</td>
			<td style="width:219px;">Asylum Research, Santa Barbara (CA), USA</td>
			<td style="width:97px;">Bioheater</td>
			<td style="width:241px;">Sample heater for AFM measurements</td>
		</tr>
		<tr height="34">
			<td height="34" style="height:34px;width:293px;">Biolever mini cantilever,&#160; BL-AC40TS-C2</td>
			<td style="width:219px;">Olympus Germany LLC, Germany</td>
			<td style="width:97px;">&#160;BL-AC40TS-C2</td>
			<td style="width:241px;">Prefered cantilever for AFM measurements</td>
		</tr>
		<tr height="17">
			<td height="17" style="height:17px;width:293px;">WSxM 5.0 Develop 6.5 (2013)</td>
			<td style="width:219px;">Nanotec Electronica S.L. , Spain</td>
			<td style="width:97px;">freeware</td>
			<td style="width:241px;">Software for AFM analysis</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:293px;">Name</td>
			<td style="width:219px;">Company</td>
			<td style="width:97px;">Catalog Number</td>
			<td style="width:241px;">Comments</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:293px;">Detergents and other equiment</td>
			<td style="width:219px;"></td>
			<td style="width:97px;"></td>
			<td style="width:241px;"></td>
		</tr>
		<tr height="19">
			<td height="19" style="height:19px;width:293px;">Calcium chloride Dihydrate (CaCl2 &#8729; 2H2O)</td>
			<td style="width:219px;">Merck KGaA</td>
			<td style="width:97px;">1.02382</td>
			<td style="width:241px;">---</td>
		</tr>
		<tr height="51">
			<td height="51" style="height:51px;width:293px;">acidic acid, 100 %, p.A.</td>
			<td style="width:219px;">CARL ROTH GmbH+CO.KG</td>
			<td style="width:97px;">3738.5</td>
			<td style="width:241px;">Danger, flammable and corrosive liquid and vapour. Causes severe skin burns and eye damage.</td>
		</tr>
		<tr height="17">
			<td height="17" style="height:17px;width:293px;">Antifoam 204</td>
			<td style="width:219px;">Sigma-Aldrich Co. LLC.</td>
			<td style="width:97px;">A6426</td>
			<td style="width:241px;">For foam suppression</td>
		</tr>
		<tr height="17">
			<td height="17" style="height:17px;width:293px;">bromophenol blue, sodium salt</td>
			<td style="width:219px;">Sigma-Aldrich Co. LLC.</td>
			<td style="width:97px;">B5525</td>
			<td style="width:241px;">---</td>
		</tr>
		<tr height="19">
			<td height="19" style="height:19px;width:293px;">Coomassie Brilliant Blue R (C45H44N3NaO7S2)</td>
			<td style="width:219px;">CARL ROTH GmbH+CO.KG</td>
			<td style="width:97px;">3862.1</td>
			<td style="width:241px;">---</td>
		</tr>
		<tr height="34">
			<td height="34" style="height:34px;width:293px;">Deoxyribonuclease II from porcine spleen</td>
			<td style="width:219px;">Sigma-Aldrich Co. LLC.</td>
			<td style="width:97px;">D4138</td>
			<td style="width:241px;">Typ IV , 2,000-6,000&#160;Kunitz units/mg protein</td>
		</tr>
		<tr height="17">
			<td height="17" style="height:17px;width:293px;">Ethanol, 95%</td>
			<td style="width:219px;">VWR, Germany</td>
			<td style="width:97px;">20827.467</td>
			<td style="width:241px;">Danger, flammable</td>
		</tr>
		<tr height="17">
			<td height="17" style="height:17px;width:293px;">glycerine, p.A.</td>
			<td style="width:219px;">CARL ROTH GmbH+CO.KG</td>
			<td style="width:97px;">3783.1</td>
			<td style="width:241px;">---</td>
		</tr>
		<tr height="19">
			<td height="19" style="height:19px;width:293px;">Gold(III) chloride trihydrate (HAuCl4 &#8729; 3H2O)</td>
			<td style="width:219px;">Sigma-Aldrich Co. LLC.</td>
			<td style="width:97px;">520918</td>
			<td style="width:241px;">Danger</td>
		</tr>
		<tr height="17">
			<td height="17" style="height:17px;width:293px;">Guanidine hydrochloride (GuHCl)</td>
			<td style="width:219px;">CARL ROTH GmbH+CO.KG</td>
			<td style="width:97px;">0037.1</td>
			<td style="width:241px;">---</td>
		</tr>
		<tr height="34">
			<td height="34" style="height:34px;width:293px;">Hellmanex III</td>
			<td style="width:219px;">Hellma GmbH &#38; Co. KG</td>
			<td style="width:97px;">9-307-011-4-507</td>
			<td style="width:241px;">---</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:293px;">Hydrochloric acid (HCl) (37%)</td>
			<td style="width:219px;">CARL ROTH GmbH+CO.KG</td>
			<td style="width:97px;">4625.2</td>
			<td style="width:241px;">Danger; Corrosive, used for pH adjustment</td>
		</tr>
		<tr height="34">
			<td height="34" style="height:34px;width:293px;">Lysozyme from chicken egg white</td>
			<td style="width:219px;">Sigma-Aldrich Co. LLC.</td>
			<td style="width:97px;">L6876&#160;</td>
			<td style="width:241px;">Lyophilized powder, protein =90 %, =40,000 units/mg protein (Sigma)&#160;</td>
		</tr>
		<tr height="19">
			<td height="19" style="height:19px;width:293px;">Magnesium chloride Hexahydrate (MgCl2 &#8729; 6H2O)</td>
			<td style="width:219px;">Merck KGaA</td>
			<td style="width:97px;">1.05833</td>
			<td style="width:241px;">---</td>
		</tr>
		<tr height="34">
			<td height="34" style="height:34px;width:293px;">Magnetic stirrer with heating,&#160; MR 3000K</td>
			<td style="width:219px;">Heidolph Instruments GmbH &#38; Co.KG, Germany</td>
			<td style="width:97px;">504.10100.00</td>
			<td style="width:241px;">Standard stirrer within experiment</td>
		</tr>
		<tr height="17">
			<td height="17" style="height:17px;width:293px;">NB-Media DM180</td>
			<td style="width:219px;">Mast Diagnostica GmbH</td>
			<td style="width:97px;">121800</td>
			<td style="width:241px;">---</td>
		</tr>
		<tr height="19">
			<td height="19" style="height:19px;width:293px;">Nitric acid (HNO3)</td>
			<td style="width:219px;">CARL ROTH GmbH+CO.KG</td>
			<td style="width:97px;">HN50.1</td>
			<td style="width:241px;">Danger; Oxidizing, Corrosing</td>
		</tr>
		<tr height="17">
			<td height="17" style="height:17px;width:293px;">PageRuler Unstained Protein Ladder</td>
			<td style="width:219px;">ThermoScientific-Pierce</td>
			<td style="width:97px;">26614</td>
			<td style="width:241px;">---</td>
		</tr>
		<tr height="17">
			<td height="17" style="height:17px;width:293px;">Poly(sodium 4-styrenesulfonat) (PSS)</td>
			<td style="width:219px;">Sigma-Aldrich Co. LLC.</td>
			<td style="width:97px;">243051</td>
			<td style="width:241px;">Average Mw ~70,000</td>
		</tr>
		<tr height="34">
			<td height="34" style="height:34px;width:293px;">Polyethylenimine (PEI), branched</td>
			<td style="width:219px;">Sigma-Aldrich Co. LLC.</td>
			<td style="width:97px;">408727</td>
			<td style="width:241px;">Warning; Harmful, Irritant, Dangerous for the environment; average Mw ~25,000</td>
		</tr>
		<tr height="19">
			<td height="19" style="height:19px;width:293px;">Potassium carbonate anhydrous (K2CO3)</td>
			<td style="width:219px;">Sigma-Aldrich Co. LLC.</td>
			<td style="width:97px;">60108</td>
			<td style="width:241px;">Warning; Harmful</td>
		</tr>
		<tr height="17">
			<td height="17" style="height:17px;width:293px;">Ribonuclease A from bovine pancreas&#160;</td>
			<td style="width:219px;">Sigma-Aldrich Co. LLC.</td>
			<td style="width:97px;">R5503</td>
			<td style="width:241px;">Type I-AS, 50-100 Kunitz units/mg protein&#160;</td>
		</tr>
		<tr height="34">
			<td height="34" style="height:34px;width:293px;">Sodium azide (NaN3)</td>
			<td style="width:219px;">Merck KGaA</td>
			<td style="width:97px;">106688</td>
			<td style="width:241px;">Danger; very toxic and Dangerous for the environment</td>
		</tr>
		<tr height="17">
			<td height="17" style="height:17px;width:293px;">Sodium chloride (NaCl)</td>
			<td style="width:219px;">CARL ROTH GmbH+CO.KG</td>
			<td style="width:97px;">3957.2</td>
			<td style="width:241px;">---</td>
		</tr>
		<tr height="17">
			<td height="17" style="height:17px;width:293px;">Sodium dodecyl sulfate (SDS)</td>
			<td style="width:219px;">Sigma-Aldrich Co. LLC.</td>
			<td style="width:97px;">L-5750</td>
			<td style="width:241px;">Danger; toxic</td>
		</tr>
		<tr height="34">
			<td height="34" style="height:34px;width:293px;">Sodium hydroxide (NaOH)</td>
			<td style="width:219px;">CARL ROTH GmbH+CO.KG</td>
			<td style="width:97px;">6771.1</td>
			<td style="width:241px;">Danger; Corrosive, used for pH regulation within cultivation and pH adjustment</td>
		</tr>
		<tr height="17">
			<td height="17" style="height:17px;width:293px;">Spectra/Por 6, Dialysis membrane, MWCO 50,000&#160;</td>
			<td style="width:219px;">CARL ROTH GmbH+CO.KG</td>
			<td style="width:97px;">1893.1</td>
			<td style="width:241px;">---</td>
		</tr>
		<tr height="34">
			<td height="34" style="height:34px;width:293px;">Sulfuric acid (H2SO4)</td>
			<td style="width:219px;">CARL ROTH GmbH+CO.KG</td>
			<td style="width:97px;">HN52.2</td>
			<td style="width:241px;">Danger; Corrosive, used for pH regulation within cultivation</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:293px;">Tannic acid (C76H52O46)</td>
			<td style="width:219px;">Sigma-Aldrich Co. LLC.</td>
			<td style="width:97px;">16201</td>
			<td style="width:241px;">---</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:293px;">TRIS HCl (C4H11NO3HCl)</td>
			<td style="width:219px;">CARL ROTH GmbH+CO.KG</td>
			<td style="width:97px;">9090.2</td>
			<td style="width:241px;">---</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:293px;">Tri-sodium citrate dihydrate (C6H5Na3O7 &#8729; 2H2O)</td>
			<td style="width:219px;">CARL ROTH GmbH+CO.KG</td>
			<td style="width:97px;">3580.2</td>
			<td style="width:241px;">---</td>
		</tr>
		<tr height="34">
			<td height="34" style="height:34px;width:293px;">Triton X-100</td>
			<td style="width:219px;">CARL ROTH GmbH+CO.KG</td>
			<td style="width:97px;">3051.3</td>
			<td style="width:241px;">Warning; Harmful, Dangerous for the environment</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">VIVASPIN 500, 50.000 MWCO Ultrafiltration tubes</td>
			<td>Sartorius AG</td>
			<td>VS0132</td>
			<td style="width:241px;">---</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:293px;">&#946;-mercaptoethanol</td>
			<td style="width:219px;">Sigma-Aldrich Co. LLC.</td>
			<td style="width:97px;">M6250</td>
			<td style="width:241px;">Danger, toxic</td>
		</tr>
	</tbody>
</table>

au-interaction of slp1 polymers and monolayer from lysinibacillus sphaericus jg-b53 - qcm-d, icp-ms and afm as tools for biomolecule-metal studies

In this publication the gold sorption behavior of surface layer (S-layer) proteins (Slp1) of Lysinibacillus sphaericus JG-B53 is described. These biomolecules arrange in paracrystalline two-dimensional arrays on surfaces, bind metals, and are thus interesting for several biotechnical applications, such as biosorptive materials for the removal or recovery of different elements from the environment and industrial processes. The deposition of Au(0) nanoparticles on S-layers, either by S-layer directed synthesis 1 or adsorption of nanoparticles, opens new possibilities for diverse sensory applications. Although numerous studies have described the biosorptive properties of S-layers 2-5, a deeper understanding of protein-protein and protein-metal interaction still remains challenging. In the following study, inductively coupled mass spectrometry (ICP-MS) was used for the detection of metal sorption by suspended S-layers. This was correlated to measurements of quartz crystal microbalance with dissipation monitoring (QCM-D), which allows the online detection of proteinaceous monolayer formation and metal deposition, and thus, a more detailed understanding on metal binding.
The ICP-MS results indicated that the binding of Au(III) to the suspended S-layer polymers is pH dependent. The maximum binding of Au(III) was obtained at pH 4.0. The QCM-D investigations enabled the detection of Au(III) sorption as well as the deposition of Au(0)-NPs in real-time during the in situ experiments. Further, this method allowed studying the influence of metal binding on the protein lattice stability of Slp1. Structural properties and protein layer stability could be visualized directly after QCM-D experiment using atomic force microscopy (AFM). In conclusion, the combination of these different methods provides a deeper understanding of metal binding by bacterial S-layer proteins in suspension or as monolayers on either bacterial cells or recrystallized surfaces.

微生物およびSLP1キャラの育成 細菌増殖の記録されたデータは、約5時間で指数増殖期の終わりを示します。以前の研究は、それにもかかわらず、定義されたメディアコンポーネントを使用して、培養の最適化またはfed-。最大収率19 4.36グラム/ L（SLP1は収穫のこの時点から単離することができることを?...

Watch this Scientific Journal Video about Au-Interaction of Slp1 Polymers and Monolayer from Lysinibacillus sphaericus JG-B53 - QCM-D, ICP-MS and AFM as Tools for Biomolecule-metal Studies at JoVE.com

Au-Interaction of Slp1 Polymers and Monolayer from Lysinibacillus sphaericus JG-B53 - QCM-D, ICP-MS and AFM as Tools for Biomolecule-metal Studies

To obtain basic information on the sorption and recycling of gold from aqueous systems the interaction of Au(III) and Au(0) nanoparticles on S-layer proteins were investigated. The sorption of protein polymers was investigated by ICP-MS and that of proteinaceous monolayers by QCM-D. Subsequent AFM enables the imaging of the nanostructures.

SLP1ポリマーと単層のAuのインタラクションから<em&gt; Lysinibacillusスフェリカス</em&gt; JG-B53 - 生体分子金属研究のためのツールと​​して、QCM-D、ICP-MSおよびAFM

In this publication the gold sorption behavior of surface layer (S-layer) proteins (Slp1) of Lysinibacillus sphaericus JG-B53 is described. These ...

au-interaction-slp1-polymers-monolayer-from-lysinibacillus-sphaericus

Universidad San Sebastian

Research

JoVE Journal

Chemistry

Au-Interaction of Slp1 Polymers and Monolayer from Lysinibacillus sphaericus JG-B53 - QCM-D, ICP-MS and AFM as Tools for Biomolecule-metal Studies

11.3K ビュー.  Helmholtz-Zentrum Dresden-Rossendorf. To obtain basic information on the sorption and recycling of gold from aqueous systems the interaction of Au(III) and Au(0) nanoparticles on S-layer proteins were investigated. The sorption of protein polymers was investigated by ICP-MS and that of proteinaceous monolayers by QCM-D. Subsequent AFM enables the imaging of the nanostructures.

ビデオ: Au-Interaction of Slp1 Polymers and Monolayer from Lysinibacillus sphaericus JG-B53 - QCM-D, ICP-MS and AFM as Tools for Biomolecule-metal Studies

Covalent Binding of BMP-2 on Surfaces Using a Self-assembled Monolayer Approach

Bone morphogenetic protein 2 (BMP-2) is a growth factor embedded in the extracellular matrix of bone tissue. BMP-2 acts as trigger of mesenchymal cell differentiation into osteoblasts, thus stimulating healing and de novo bone formation. The clinical use of recombinant human BMP-2 (rhBMP-2) in conjunction with scaffolds has raised recent controversies, based on the mode of presentation and the amount to be delivered. The protocol presented here provides a simple and efficient way to deliver BMP-2 for in vitro studies on cells. We describe how to form a self-assembled monolayer consisting of a heterobifunctional linker, and show the subsequent binding step to obtain covalent immobilization of rhBMP-2. With this approach it is possible to achieve a sustained presentation of BMP-2 while maintaining the biological activity of the protein. In fact, the surface immobilization of BMP-2 allows targeted investigations by preventing unspecific adsorption, while reducing the amount of growth factor and, most notably, hindering uncontrolled release from the surface. Both short- and long-term signaling events triggered by BMP-2 are taking place when cells are exposed to surfaces presenting covalently immobilized rhBMP-2, making this approach suitable for in vitro studies on cell responses to BMP-2 stimulation.

We describe a method for accomplishing efficient immobilization of BMP-2 on surfaces. Our approach is based on the formation of a self-assembled monolayer to achieve the covalent binding of BMP-2 via its free amine residues. This method is a useful tool to study signaling at the cell membrane.

共有結合は、自己組織化単分子膜の手法を用いて表面上のBMP-2の結合

Bone morphogenetic protein 2 (BMP-2) is a growth factor embedded in the extracellular matrix of bone tissue. BMP-2 acts as trigger of mesenchymal cell ...

化学

Manufacturing of Three-dimensionally Microstructured Nanocomposites through Microfluidic Infiltration

Microstructured composite beams reinforced with complex three-dimensionally (3D) patterned nanocomposite microfilaments are fabricated via nanocomposite in&#64257;ltration of 3D interconnected microfluidic networks. The manufacturing of the reinforced beams begins with the fabrication of microfluidic networks, which involves layer-by-layer deposition of fugitive ink filaments using a dispensing robot, filling the empty space between filaments using a low viscosity resin, curing the resin and finally removing the ink.&#160;Self-supported 3D structures with other geometries and many layers (e.g.&#160;a few hundreds layers) could be built using this method. The resulting tubular micro&#64258;uidic networks are then infiltrated with thermosetting nanocomposite suspensions containing nanofillers (e.g.&#160;single-walled carbon nanotubes), and subsequently cured. The infiltration is done by applying a pressure gradient between two ends of the empty network (either by applying a vacuum or vacuum-assisted microinjection). Prior to the infiltration, the nanocomposite suspensions are prepared by dispersing nanofillers into polymer matrices using ultrasonication and three-roll mixing methods. The nanocomposites (i.e.&#160;materials infiltrated) are then solidified under UV exposure/heat cure, resulting in a 3D-reinforced composite structure. The technique presented here enables the design of functional nanocomposite macroscopic products for microengineering applications such as actuators and sensors.

Three-dimensional (3D) microstructured composite beams are fabricated through the directed and localized infiltration of nanocomposites into 3D porous microfluidic networks. The flexibility of this manufacturing method enables the utilization of different thermosetting materials and nanofillers in order to achieve a variety of functional 3D reinforced nanocomposite macroscopic products.

マイクロ流体浸透を通じて立体的微細構造ナノ複合材料の製造

Microstructured composite beams reinforced with complex three-dimensionally (3D) patterned nanocomposite microfilaments are fabricated via nanocomposite ...

Standards for Quantitative Metalloproteomic Analysis Using Size Exclusion ICP-MS

Metals are essential for protein function as cofactors to catalyze chemical reactions. Disruption of metal homeostasis is implicated in a number of diseases including Alzheimer's and Parkinson's disease, but the exact role these metals play is yet to be fully elucidated. Identification of metalloproteins encounters many challenges and difficulties. Here we report an approach that allows metalloproteins in complex samples to be quantified. This is achieved using size exclusion chromatography coupled with inductively coupled plasma - mass spectrometry (SEC-ICP-MS). Using six known metalloproteins, the size exclusion column can be calibrated and the respective trace elements (iron, copper, zinc, cobalt, iodine) can be used for quantification. SEC-ICP-MS traces of human brain and plasma are presented. The use of these metalloprotein standards provides the means to quantitatively compare metalloprotein abundances between biological samples. This technique is poised to help shed light on the role of metalloproteins in neurodegenerative disease as well as other diseases where imbalances in trace elements are implicated.

Size exclusion chromatography hyphenated with inductively coupled plasma - mass spectrometry (ICP-MS) is a powerful tool to measure changes in the abundance of metalloproteins directly from biological samples. Here we describe a set of metalloprotein standards used to estimate molecular mass and the amount of metal associated with unknown proteins.

サイズ排除ICP-MSを用いた定量Metalloproteomic分析のための基準

Metals are essential for protein function as cofactors to catalyze chemical reactions. Disruption of metal homeostasis is implicated in a number of ...

HKUST-1 as a Heterogeneous Catalyst for the Synthesis of Vanillin

Vanillin (4-hydoxy-3-methoxybenzaldehyde) is the main component of the extract of vanilla bean. The natural vanilla scent is a mixture of approximately 200 different odorant compounds in addition to vanillin. The natural extraction of vanillin (from the orchid Vanilla planifolia, Vanilla tahitiensis and Vanilla pompon) represents only 1% of the worldwide production and since this process is expensive and very long, the rest of the production of vanillin is synthesized. Many biotechnological approaches can be used for the synthesis of vanillin from lignin, phenolic stilbenes, isoeugenol, eugenol, guaicol, etc., with the disadvantage of harming the environment since these processes use strong oxidizing agents and toxic solvents. Thus, eco-friendly alternatives on the production of vanillin are very desirable and thus, under current investigation. Porous coordination polymers (PCPs) are a new class of highly crystalline materials that recently have been used for catalysis. HKUST-1 (Cu3(BTC)2(H2O)3, BTC = 1,3,5-benzene-tricarboxylate) is a very well known PCP which has been extensively studied as a heterogeneous catalyst. Here, we report a synthetic strategy for the production of vanillin by the oxidation of trans-ferulic acid using HKUST-1 as a catalyst.

The conversion of trans-ferulic acid to vanillin was achieved by heterogeneous catalysis. HKUST-1 was employed in this synthesis and the essential step in the catalytic process was the generation of unsaturated metal sites. Thus, when the catalyst was activated under vacuum, full vanillin conversion (yield of 95%) was obtained.

HKUST-1バニリンの合成のための不均一触媒として、

Vanillin (4-hydoxy-3-methoxybenzaldehyde) is the main component of the extract of vanilla bean. The natural vanilla scent is a mixture of approximately ...

Electrochemical Impedance Spectroscopy as a Tool for Electrochemical Rate Constant Estimation

Electrochemical impedance spectroscopy (EIS) was used for advanced characterization of organic electroactive compounds along with cyclic voltammetry (CV). In the case of fast reversible electrochemical processes, current is predominantly affected by the rate of diffusion, which is the slowest and limiting stage. EIS is a powerful technique that allows separate analysis of stages of charge transfer that have different AC frequency response. The capability of the method was used to extract the value of charge transfer resistance, which characterizes the rate of charge exchange on the electrode-solution interface. The application of this technique is broad, from biochemistry up to organic electronics. In this work, we are presenting the method of analysis of organic compounds for optoelectronic applications.

Electrochemical impedance spectroscopy (EIS) of species that undergo reversible oxidation or reduction in solution was used for determination of rate constants of oxidation or reduction.

電気化学インピー ダンス分光電気化学定数推定法のためのツールとして

Electrochemical impedance spectroscopy (EIS) was used for advanced characterization of organic electroactive compounds along with cyclic voltammetry (CV). ...

Raman and IR Spectroelectrochemical Methods as Tools to Analyze Conjugated Organic Compounds

In the presented work, two spectroelectrochemical techniques are discussed as tools for the analysis of the structural changes occurring in the molecule on the vibrational level of energy. Raman and IR spectroelectrochemistry can be used for advanced characterization of the structural changes in the organic electroactive compounds. Here, the step-by-step analysis by means of Raman and IR spectroelectrochemistry is shown. Raman and IR spectroelectrochemical techniques provide complementary information about structural changes occurring during an electrochemical process, i.e. allows for the investigation of redox processes and their products. The examples of IR and Raman spectroelectrochemical analysis are presented, in which the products of the redox reactions, both in solution and solid state, are identified.

A protocol of step-by-step Raman and IR spectroelectrochemical analysis is presented.

ラマン散乱とツールとしての IR 分光電気化学法共役有機化合物を分析するには

In the presented work, two spectroelectrochemical techniques are discussed as tools for the analysis of the structural changes occurring in the molecule ...

Using Cyclic Voltammetry, UV-Vis-NIR, and EPR Spectroelectrochemistry to Analyze Organic Compounds

Cyclic voltammetry (CV) is a technique used in the analysis of organic compounds. When this technique is combined with electron paramagnetic resonance (EPR) or ultraviolet-visible and near-infrared (UV-Vis-NIR) spectroscopies, we obtain useful information such as electron affinity, ionization potential, band-gap energies, the type of charge carriers, and degradation information that can be used to synthesize stable organic electronic devices. In this study, we present electrochemical and spectroelectrochemical methods to analyze the processes occurring in active layers of an organic device as well as the generated charge carriers.

In this article, we describe electrochemical, electron paramagnetic resonance, and ultraviolet-visible and near-infrared spectroelectrochemical methods to analyze organic compounds for application in organic electronics.

サイクリックボルタンメトリー、紫外-可視-近赤外、および EPR 分光電気化学を使用して有機性化合物を分析するには

Cyclic voltammetry (CV) is a technique used in the analysis of organic compounds. When this technique is combined with electron paramagnetic resonance ...

Preparation of Stable Bicyclic Aziridinium Ions and Their Ring-Opening for the Synthesis of Azaheterocycles

Bicyclic aziridinium ions were generated by the removal of an appropriate leaving group through internal nucleophilic attack by nitrogen atom in the aziridine ring. The utility of bicyclic aziridinium ions, specifically 1-azoniabicyclo[3.1.0]hexane and 1-azoniabicyclo[4.1.0]heptane tosylate highlighted in the aziridine ring openings by the nucleophile with the release of the ring strain to yield the corresponding ring-expanded azaheterocycles such as pyrrolidine, piperidine and azepane with diverse substituents on the ring in regio- and stereospecific manner. Herein, we report a simple and convenient method for the preparation of the stable 1-azabicyclo[4.1.0]heptane tosylate followed by selective ring opening via a nucleophilic attack either at the bridge or at the bridgehead carbon to yield piperidine and azepane rings, respectively. This synthetic strategy allowed us to prepare biologically active natural products containing piperidine and azepane motif including sedamine, allosedamine, fagomine and balanol in highly efficient manner.

Bicyclic aziridinium ions such as 1-azoniabicyclo[4.1.0]heptane tosylate were generated from 2-[4-tolenesulfonyloxybutyl]aziridine, which was utilized for the preparation of substituted piperidines and azepanes via regio- and stereospecific ring-expansion with various nucleophiles. This highly efficient protocol allowed us to prepare diverse azaheterocycles including natural products such as fagomine, febrifugine analogue and balanol.

安定した二環性 Aziridinium イオンと開環の Azaheterocycles の合成のための準備

Bicyclic aziridinium ions were generated by the removal of an appropriate leaving group through internal nucleophilic attack by nitrogen atom in the ...

Stereolithographic 3D Printing with Renewable Acrylates

The accessibility of cost-competitive renewable materials and their application in additive manufacturing is essential for an efficient biobased economy. We demonstrate the rapid prototyping of sustainable resins using a stereolithographic 3D printer. Resin formulation takes place by straightforward mixing of biobased acrylate monomers and oligomers with a photoinitiatior and optical absorber. Resin viscosity is controlled by the monomer to oligomer ratio and is determined as a function of shear rate by a rheometer with parallel plate geometry. A stereolithographic apparatus charged with the biobased resins is employed to produce complex shaped prototypes with high accuracy. The products require a post-treatment, including alcohol rinsing and UV irradiation, to ensure complete curing. The high feature resolution and excellent surface finishing of the prototypes is revealed by scanning electron microscopy.

A protocol for additive manufacturing with renewable photopolymer resins on a stereolithography apparatus is presented.

再生可能なアクリレートと Stereolithographic 3 D プリント

The accessibility of cost-competitive renewable materials and their application in additive manufacturing is essential for an efficient biobased economy. ...

A Continuous-flow Photocatalytic Reactor for the Precisely Controlled Deposition of Metallic Nanoparticles

In this work, a novel photocatalytic reactor for the pulsed and controlled excitation of the photocatalyst and the precise deposition of metallic nanoparticles is developed. Guidelines for the replication of the reactor and its operation are provided in detail. Three different composite systems (Pt/graphene, Pt/TiO2, and Au/TiO2) with monodisperse and uniformly distributed particles are produced by this reactor, and the photodeposition mechanism, as well as the synthesis optimization strategy, are discussed. The synthesis methods and their technical aspects are described comprehensively. The role of the ultraviolet (UV) dose (in each excitation pulse) on the photodeposition process is investigated and the optimum values for each composite system are provided.

For a continuous and scalable synthesis of noble-metal-based nanocomposites, a novel photocatalytic reactor is developed and its structure, operation principles, and product quality optimization strategies are described.